User Manual

User Manual
TC-5000
OPERATOR'S MANUAL
Issue 1
5/09
CONTENTS
PAGE
INTRODUCTION
3
SAFETY AND INSTALLATION INFORMATION
Operator Safety
Warning
Unpacking
Installation
Replacement Cable
Working Conditions
Guarantee
4
4
4
4
4
5
5
6
SÉCURITÉ ET CONSIGNES D'INSTALLATION
Sécurité de l'opérateur
Avertissement
Déballage
Installation
Câble de rechange
Conditions de travail
Garantie
7
7
7
7
7
8
8
9
SICHERHEITS - UND INSTALLATIONSINFORMATIONEN
Sicherheit des Bedienpersonals
Achtung
Auspacken
Inbetriebnahme
Ersatzkabel
Einsatzbedingungen
Garantie
10
10
10
10
10
11
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12
INFORMACIÓN DE SEGURIDAD E INSTALACIÓN
Seguridad del operario
Advertencia
Desembalaje
Instalación
Cable de repuesto
Condiciones de trabajo
Garantía
13
13
13
13
13
14
14
15
INFORMAZIONI SULLA SICUREZZA E L'INSTALLAZIONE
Sicurezza dell'operatore
Avvertenza
Disimballaggio
Installazione
Cavo di ricambio
Condizioni di esercizio
Garanzia
16
16
16
16
16
17
17
18
1
INSTRUMENT CONNECTIONS
19
CONTACT INFORMATION
20
TC-5000 SPECIFICATION
21
TC-5000 OPERATION AND MAINTENANCE
Gradient
Adjustable Heated Lid
Adjustable Screen
Consumables
PC Connections
Cleaning the TC-5000
Removing the Block
23
23
24
25
25
26
27
28
PROGRAMMING THE TC-5000
Start up Menu
The System User Menu
Writing and Editing Programs
Option 1: Copying an Existing Template
Option 2: Editing and Saving an Existing Template
Option 3: Creating a New Program
Inserting a Segment
Inserting Cycle Numbers
Inserting Incremental or Decremental Time or Temperature
Inserting a Pause
Saving Changes to a Program
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29
30
31
32
34
35
36
37
37
39
39
RUNNING A PROGRAM
Program Defaults
Gradient Calculator
39
42
44
SYSTEM ADMINISTRATION
Program Passwords
System Administration Password
System Test
Controller Reset
System Configuration
45
46
46
47
47
48
FREQUENTLY ASKED QUESTIONS
50
ADDITIONAL INFORMATION
Fault Finding
Fuses
Accessories
PC Accessories
Replacement Parts
51
51
51
51
52
52
2
INTRODUCTION
The TC-5000 is a gradient thermal cycler, designed to accurately control the temperatures of samples
between 4°C and 99°C. It has many scientific applications including DNA amplification, PCR 1, reverse
transcription and sequencing.
The TC-5000 is programmed via a touch screen LCD graphics display. The user is able to input various
programs and parameters, including:
•
•
•
•
•
Specified temperatures in °C
Times for which the sample is to be held at the specified temperature
Heating and cooling rates between the specified temperatures in °C/sec
Gradient of temperatures across the thermal cycling block
Incremental time or temperature increases or decreases when a cycle is repeated
Before using the TC-5000 make sure you have read this manual carefully. If there is any doubt relating
to the proper use of this equipment, the staff at Bibby Scientific Ltd or your supplier will be happy to
assist you.
TC
-5
00
0
1
Purchase of this instrument conveys a limited non-transferable immunity from suit for the purchaser's own internal research and
development and applied fields other than human in-vitro diagnostics under non-real-time thermal cycler patents of Applied Biosystems LLC.
3
SAFETY AND INSTALLATION
OPERATOR SAFETY
Please read this manual carefully before using the thermal cycler. If the equipment is not used in the
manner described in this manual the protection provided by the equipment might be impaired.
All Bibby Scientific Ltd instruments are designed to conform to international safety requirements and
are fitted with an over-temperature cut-out. If a safety problem should be encountered then switch off
the unit at the mains socket and remove the plug from the electricity supply.
WARNING
HIGH TEMPERATURES ARE DANGEROUS as they can cause serious burns to operators and ignite
combustible material. Users should be aware of the following potential hazards:
•
•
•
•
USE CARE AND WEAR PROTECTIVE GLOVES TO PROTECT HANDS
DO NOT use combustible substances near hot objects
DO NOT operate the instrument in the vicinity of inflammable liquids or gases
DO NOT place any liquid directly into the instrument, always use the appropriate
plastic consumables
After heating samples, remember that parts of the instrument, namely the lid and block and any
associated accessories may be very hot.
UNPACKING
When unpacking please ensure that the following have been removed from the packaging:
•
•
•
•
Thermal cycler
Mains cable
Guarantee card
Decontamination certificate
The user is advised to keep the original packaging in case the instrument ever needs to be returned for
service or repair. Bibby Scientific Ltd accepts no responsibility for damage incurred unless the unit is
correctly packed and transported in its original packaging.
Please note that the screen of the thermal cycler is easily damaged by sharp objects such as pens,
pencils and fingernails. This type of damage will be considered as misuse and invalidate the guarantee
for this component.
INSTALLATION
The instrument should be carried using both hands. Never move or carry the instrument when in use or
connected to the mains electricity supply.
1. All Bibby Scientific Ltd instruments are supplied with a power cable; this may be integral or
plug-in.
2. Before connecting the instrument to the mains electricity supply, check the voltage against the
4
rating plate (located on the back of the unit). Ensure that the voltage selector switch (located above
the On/Off switch) is set to the appropriate voltage for the local supply. Please note that the unit
must be earthed to ensure proper electrical safety. Connect the mains cable to a suitable plug
according to the table below.
Connections
Live
Neutral
Earth
220V-240V, 50/60Hz Supply
brown
blue
green/yellow
110V-120V, 50/60Hz Supply
black
white
green
3. UK ONLY: The fused plug supplied with the mains cable is fitted with a 10 Amp fuse to protect the
instrument and the user.
4. Units showing 230V, 50/60Hz on the rating plate also operate between 210 and 260V; units with
120V, 50/60Hz also operate between 100 and 130V and units marked with 100V will operate
between 90 and 110V. In all cases the heating rate will degrade by approximately 8% at the
extremes of the voltage range.
5. Plug the mains cable into the socket on the back of the instrument.
6. Place the unit on a suitable flat bench or in a fume cupboard if required, ensuring that the air inlet
vents on the underside are free from obstruction. Position the instrument with a minimum distance
all round of 200mm from walls or other items and between each unit if multiple units are being
used to permit efficient air flow for each instrument.
7. Switch on the instrument:
I mains switch On
O mains switch Off
REPLACEMENT CABLE
Should the mains lead need replacement, a cable of 1mm2 of harmonized code H05VV-F connected to
an IEC320 plug should be used. IF IN DOUBT CONSULT A QUALIFIED ELECTRICIAN.
WORKING CONDITIONS
The thermal cycler is designed to operate under the following conditions:
•
•
•
•
•
•
Indoor use
Ambient temperature range +5°C to +40°C
Altitude to 2000m
Mains supply fluctuations not exceeding 10%
Over voltage category II IEC 60364-4-443
Pollution degree 2
Note: The control specifications are quoted at an ambient temperature of 20°C. The specification may
deteriorate outside an ambient temperature of 10°C to 30°C.
The instrument has been tested for radio frequency interference and is certified under EN61326.
5
GUARANTEE
The instrument is guaranteed against any defect in material or workmanship for the period specified
on the enclosed guarantee card. This period is effective from the date of purchase; within this period
all defective parts will be replaced free of charge provided that the defect is not the result of an
accident, misuse or negligence. The screen of the thermal cycler can be damaged by sharp objects such
as pens, pencils and fingernails. Damage of this sort will be considered misuse and will invalidate the
guarantee for this component.
Servicing under this guarantee should be obtained from the supplier of the instrument.
This manual has been prepared for the convenience of Bibby Scientific Ltd's customers and nothing in
this manual shall be taken as a warranty, condition or representation concerning the description,
merchantability, fitness for purpose or otherwise of the unit or components.
Notwithstanding the description and specification(s) of the instruments contained in the operator's
manual, Bibby Scientific Ltd reserves the right to make such changes as it sees fit to the instruments or
to any of the components.
6
SÉCURITÉ ET CONSIGNES D'INSTALLATION
SÉCURITÉ DE L’OPÉRATEUR
Lire attentivement le présent manuel avant d'utiliser le cycleur thermique. Si ce matériel n'est pas utilisé
de la manière décrite dans le présent manuel, la protection fournie risque d'être compromise.
Tous les appareils Bibby Scientific Ltd sont conçus pour être conformes aux exigences internationales de
sécurité et sont dotés d'une coupure en cas de surchauffe. Si un problème de sécurité survient, mettre
l'appareil hors tension au niveau de la prise secteur et débrancher la fiche de l'alimentation électrique.
AVERTISSEMENT
Les TEMPÉRATURES ÉLEVÉES SONT DANGEREUSES car elles peuvent provoquer de graves brûlures chez
l'opérateur et enflammer les matériaux combustibles. Les utilisateurs devront être conscients des
dangers potentiels suivants :
•
•
•
•
AGIR AVEC PRUDENCE ET PORTER DES GANTS DE PROTECTION POUR LES MAINS
NE PAS utiliser de substances combustibles à proximité d'objets chauds
NE PAS utiliser l'appareil au voisinage de liquides ou de gaz inflammables
NE PAS placer un quelconque liquide directement sur l'appareil; toujours utiliser les
consommables en plastique appropriés
Rappel : après le chauffage d'échantillons, certaines parties de l'appareil, notamment le couvercle et le
bloc, ainsi que les accessoires associés, peuvent être brûlants.
DÉBALLAGE
Au moment du déballage, vérifier que l'emballage contenait les articles suivants :
•
•
•
•
Cycleur thermique
Câble secteur
Carte de garantie
Certificat de décontamination
Il est conseillé à l'utilisateur de conserver l'emballage d'origine s'il devait s'avérer nécessaire de renvoyer
l'appareil pour une intervention d'entretien ou de réparation. Bibby Scientific Ltd n'accepte aucune
responsabilité pour les dégâts survenus si l'appareil n'est pas correctement emballé et transporté dans
sont emballage d'origine.
Noter que des objets pointus tels que crayons, stylos et ongles peuvent facilement endommager l'écran
du cycleur thermique. Ce type de dégâts sera considéré comme une mauvaise utilisation et annulera la
garantie de ce composant.
INSTALLATION
Porter l'appareil à deux mains. Ne jamais déplacer ou transporter l'appareil lorsqu'il est en service ou
branché à l'alimentation électrique sur secteur.
1. Tous les appareils Bibby Scientific Ltd sont livrés avec un câble d'alimentation, qui peut être intégré à
l'appareil ou muni d'une fiche.
7
2. Avant de raccorder l'appareil à l'alimentation électrique sur secteur, vérifier la tension par rapport à
la plaque signalétique (située au dos de l'appareil). Vérifier que le sélecteur de tension (situé audessus de l'interrupteur Marche/Arrêt) est réglé sur la tension correspondant au réseau local. Noter
que l'appareil doit être relié à la terre pour assurer une bonne sécurité électrique. Brancher le câble
secteur sur une fiche adéquate d'après le tableau suivant.
Connexions
Phase
Neutre
Terre
Alimentation 220V-240V, 50/60Hz
marron
bleu
vert/jaune
Alimentation 110V-120V, 50/60Hz
noir
blanc
vert
3. ROYAUME-UNI SEULEMENT: La prise à fusible fournie avec le câble secteur est munie d'un fusible 10
A destiné à protéger l'appareil et l'utilisateur.
4. Les appareils portant l'indication 230V, 50/60Hz sur la plaque signalétique fonctionnent également
entre 210 et 260V; les appareils portant l'indication 120V, 50/60Hz fonctionnent également entre
100 et 130V et les appareils portant l'indication 100V fonctionneront entre 90 et 110V. Dans tous
les cas, la puissance de chauffage se dégrade d'environ 8% aux extrêmes de la plage de tension.
5. Brancher le câble secteur dans la prise au dos de l'appareil.
6. Disposer l'appareil sur une paillasse ou sous une hotte aspirante si nécessaire, en veillant à ce que les
prises d'air situées au-dessous ne soient pas obstruées. Positionner l'appareil avec tout autour un
écartement minimum de 200mm par rapport aux murs ou autres éléments, ainsi qu'entre chaque
appareil en cas d'utilisation d'appareils multiples, afin de permettre une circulation d'air efficace pour
chaque appareil.
7. Allumer l'appareil.
I Interrupteur secteur en position Marche
O Interrupteur secteur en position Arrêt
CÂBLE DE RECHANGE
S'il s'avère nécessaire de remplacer le cordon secteur, utiliser un câble de 1 mm2 au code harmonisé
H05VV-F raccordé à une fiche IEC320. EN CAS DE DOUTE, CONSULTER UN ÉLECTRICIEN QUALIFIÉ.
CONDITIONS DE TRAVAIL
Le cycleur thermique est conçu pour fonctionner dans les conditions suivantes:
•
•
•
•
•
•
Utilisation en intérieur
Plage de température ambiante : +5°C à +40°C
Altitude jusqu'à 2000m
Fluctuations de l'alimentation sur secteur ne dépassant pas 10%
Surtensions de catégorie II selon la norme IEC 60364-4-443
Degré de pollution 2
Remarque: Les caractéristiques de régulation sont indiquées à une température ambiante de 20°C. Ces
caractéristiques peuvent se détériorer en dehors d'une température ambiante de 10°C à 30°C.
8
L'appareil a été testé en matière de perturbations radioélectriques et est certifié selon la norme
EN61326.
GARANTIE
L'appareil est garanti contre tout défaut de matériaux ou vice de fabrication pendant la période
précisée sur la carte de garantie jointe. Cette période s'applique à compter de la date d'achat; pendant
cette période, toutes les pièces défectueuses seront remplacées gratuitement à condition que le défaut
ne soit pas le résultat d'un accident, d'une mauvaise utilisation ou d'une négligence. Des objets pointus
tels que crayons, stylos et ongles peuvent endommager l'écran du cycleur thermique. Les dégâts de
cette sorte seront considérés comme une mauvaise utilisation et annuleront la garantie de ce
composant.
Dans le cadre de cette garantie, le service après-vente est à obtenir auprès du fournisseur de l'appareil.
Le présent manuel a été préparé pour le confort des clients de Bibby Scientific Ltd et rien dans son
contenu ne doit être pris comme une garantie, une condition ou une affirmation concernant la
description, la qualité marchande, l'adéquation à un usage particulier ou autre de l'appareil ou de ses
composants.
Nonobstant la description et les caractéristiques techniques des appareils figurant dans le manuel de
l'utilisateur, Bibby Scientific Ltd se réserve le droit d'apporter aux appareils ou à n'importe quel
composant les changements jugés utiles.
9
SICHERHEITS - UND INSTALLATIONSINFORMATIONEN
SICHERHEIT DES BEDIENPERSONALS
Lesen Sie diese Anleitung vor Verwendung des Thermocyclers bitte sorgfältig durch. Wenn das Gerät
nicht entsprechend der Bedienungsanleitung eingesetzt wird, können die Schutzfunktionen des Gerätes
beeinträchtigt werden.
Alle Geräte von Bibby Scientific Ltd entsprechen den internationalen Sicherheitsanforderungen und sind
mit einer Übertemperatur-Schutzvorrichtung ausgestattet. Bei einer Sicherheitsstörung bitte das Gerät
an der Steckdose ausschalten und den Netzstecker ziehen.
ACHTUNG
HOHE TEMPERATUREN STELLEN EINE GEFAHRENQUELLE DAR. Sie können schwere Brandverletzung
verursachen und brennbare Stoffe entzünden. Der Benutzer sollt sich mit den möglichen Gefahren
vertraut machen:
•
•
•
•
UMSICHTIG VORGEHEN UND SCHUTZHANDSCHUHE TRAGEN
KEINE brennbaren Stoffe in der Nähe heißer Gegenstände verwenden
Das Gerät NICHT in der Nähe entzündlicher Flüssigkeiten oder Gase betreiben
Flüssigkeiten NICHT direkt auf das Gerät auftragen - verwenden Sie stets die
entsprechenden Kunststoffeinsätze
Nach der Erhitzung von Proben daran denken, dass bestimmte Geräteteile wie Deckel, Thermoblock
und Zubehörteile sehr heiß sein können.
AUSPACKEN
Beim Auspacken darauf achten, dass der folgende Lieferumfang vorhanden ist:
•
•
•
•
Thermocycler
Netzkabel
Garantiekarte
Dekontaminationszertifikat
Wir empfehlen die Originalverpackung aufzubewahren, falls das Gerät zwecks Wartung oder Reparatur
zurückgeschickt werden muss. Bibby Scientific Ltd übernimmt keine Verantwortung für
Transportschäden, wenn das Gerät nicht ordnungsgemäß verpackt in der Originalpackung verschickt
wird.
Achten Sie bitte darauf, dass die Anzeige des Thermocyclers durch spitze bzw. scharfe Gegenstände
wie Stifte, Bleistifte und Fingernägel leicht zerkratzt wird. Diese Art von Beschädigung gilt als
Missbrauch des Geräts und führt zur Ungültigkeit der Garantie für dieses Gerät.
INBETRIEBNAHME
Das Gerät mit beiden Händen tragen. Das Gerät unter keinen Umständen transportieren, wenn es in
Betrieb ist, oder während das Gerät noch am Netz angeschlossen ist.
10
1. Alle Geräte von Bibby Scientific Ltd werden mit einem Netzkabel geliefert, das entweder eingesteckt
wird oder fest mit dem Gerät verbunden ist.
2. Vor dem Anschluss bitte kontrollieren, ob die Stromversorgung den Angaben auf dem Typenschild
(auf der Geräterückseite) entspricht. Darauf achten, dass der Spannungswähler (über dem Ein/AusSchalter) auf die entsprechende örtliche Spannung eingestellt ist. Um die elektrische Sicherheit zu
gewährleisten, muss dieses Gerät geerdet werden. Schließen Sie das Netzkabel entsprechend der
folgenden Tabelle an einen geeigneten Stecker an.
Anschluss
Phase
Neutral
Erde
220/240V, 50/60Hz
Braun
Blau
Grün/Gelb
110V/120V, 50/60Hz
Schwarz
Weiß
Grün
3. NUR FÜR GROSSBRITANNIEN: der mit dem Netzkabel gelieferte Sicherungsstecker enthält eine 10
Amp. Sicherung zum Schutz des Geräts und des Anwenders.
4. Geräte mit der Bezeichnung 230V, 50/60Hz auf dem Typenschild können mit Spannungen zwischen
210 und 260V betrieben werden; Geräte für 120V, 50/60Hz können mit Spannungen zwischen 100
und 130V betrieben werden. Bei Geräten für 100V gilt der Bereich 90 bis 110V. In allen Fällen
reduziert sich die Erhitzungsrate um etwa 8% in den äußeren Spannungsbereichen.
5. Stecken Sie das Netzkabel in die Buchse auf der Geräterückseite ein.
6. Stellen Sie das Gerät auf einen geeigneten ebenen Tisch oder in einem Abzugsschrank auf und
sorgen Sie dafür, dass die Lufteinlassschlitze auf der Geräteunterseite nicht blockiert sind. Das Gerät
muss einen Mindestabstand von 200mm zu Wänden und anderen Gegenständen bzw. zu anderen
Geräten dieser Art aufweisen, um einen ausreichenden Luftstrom zu gewährleisten.
7. Schalten Sie das Gerät ein:
I
Netzschalter Ein
O Netzschalter Aus
ERSATZKABEL
Bei einem eventuellen Austausch des Netzkabels wird ein Kabel vom Typ H05VV-F mit 1 mm2
Adernquerschnitt und Europastecker (IEC 320) benötigt. IM ZWEIFELSFALL EINEN ELEKTROFACHMANN HINZUZIEHEN.
EINSATZBEDINGUNGEN
Der Thermocycler ist für den Einsatz unter folgenden Bedingungen ausgelegt:
•
•
•
•
•
•
•
Gebrauch in Innenräumen
Umgebungstemperatur zwischen +5°C to +40°C
Höhe: bis zu 2000 m
Relative Feuchte nicht über 95%
Netzspannungsschwankungen nicht über 10%
Überspannungsklasse 2 IEC 60364-4-443
Verschmutzungsgrad 2
11
Hinweis: Die Gerätespezifikationen beziehen sich auf eine Umgebungstemperatur von 20°C und
können sich außerhalb des Bereichs 10°C bis 30°C verschlechtern.
Das Gerät wurde auf HF-Störeinflüsse geprüft und entspricht den EMV-Bedingungen nach EN61326.
GARANTIE
Bibby Scientific Ltd gewährleistet, dass dieses Gerät für den auf der Garantiekarte angegebenen
Zeitraum keine Herstellungs- und Materialmängel aufweist. Dieser Zeitraum tritt ab dem Verkaufsdatum
in Kraft. Innerhalb dieses Zeitraums werden alle defekten Teile kostenlos ausgetauscht, soweit der
Defekt nicht auf einen Unfall, Missbrauch oder Nachlässigkeit zurückzuführen ist. Die Anzeige des
Thermocylers kan durch spitze/scharfe Gegenstände wie Stifte, Bleistifte und Fingernägel beschädigt
werden. Diese Art von Beschädigung gilt als Missbrauch des Geräts und führt zur Ungültigkeit der
Garantie für dieses Gerät.
Wartungsarbeiten, die unter diese Garantie fallen, müssen von der Verkaufsstelle für dieses Gerät
gehandhabt werden.
Diese Anleitung wurde zur Information der Kunden von Bibby Scientific Ltd erstellt und stellt in keinster
Weise eine Gewährleistung, Bedingung oder Darstellung bezüglich der Beschreibung, Marktgängigkeit
oder Zweckdienlichkeit dieser Geräte oder Bauteile dar.
Unabhängig von Beschreibung und Spezifikation(en) des hier beschriebenen Geräts behält sich Bibby
Scientific Ltd das Recht vor, Änderungen an diesem Gerät oder dessen Bauteilen vorzunehmen.
12
INFORMACIÓN DE SEGURIDAD E INSTALACIÓN
SEGURIDAD DEL OPERARIO
Lea atentamente este manual antes de utilizar el ciclador térmico. Si el equipo no se utiliza de la forma
descrita en este manual, se reducirá la protección ofrecida por el equipo.
Todos los instrumentos Bibby Scientific Ltd están diseñados para cumplir los requisitos internacionales
de seguridad, e incluyen un dispositivo de corte de sobretemperatura. Si se produce un problema de
seguridad, apague la unidad en la toma de alimentación y retire el enchufe del suministro eléctrico.
ADVERTENCIA
LAS ALTAS TEMPERATURAS SON PELIGROSAS, ya que pueden ocasionar quemaduras graves a los
operarios y prender el material combustible. Los usuarios deben conocer los posibles riesgos:
• TENGA CUIDADO Y LLEVE GUANTES DE PROTECCIÓN PARA PROTEGERSE LAS
MANOS
• NO utilice sustancias combustibles cerca de objetos calientes
• NO utilice el instrumento cerca de líquidos o gases inflamables
• NO coloque un líquido directamente en el instrumento. Utilice siempre los
consumibles plásticos adecuados
Después de calentar las muestras, recuerde que hay componentes del instrumento que pueden
calentarse mucho, concretamente la tapa y el cuerpo y todos los accesorios asociados.
DESEMBALAJE
Durante el desembalaje, asegúrese de sacar los siguientes componentes del embalaje:
•
•
•
•
Ciclador térmico
Cable de alimentación
Tarjeta de garantía
Certificado de descontaminación
Se recomienda guardar el embalaje original en caso de que tenga que enviar el instrumento para un
trabajo de mantenimiento o reparación. Bibby Scientific Ltd no se responsabiliza de los daños
producidos si la unidad no está debidamente embalada y no se envía en su embalaje original.
Tenga en cuenta que los objetos punzantes, p.ej.: bolígrafos, lápices y uñas, pueden dañar fácilmente
la pantalla del ciclador térmico. Este tipo de daño se considerará como un uso incorrecto, e invalidará
la garantía de este componente.
INSTALACIÓN
El instrumento se debe transportar con las dos manos. No mueva ni lleve el instrumento cuando se
utilice o esté conectado al suministro eléctrico principal.
1. Todos los instrumentos Bibby Scientific Ltd se suministran con un cable de alimentación, que puede
ser integrado o 'enchufable'.
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2. Antes de conectar el instrumento al suministro eléctrico, compruebe que el voltaje coincida con el
indicado en la placa de régimen (situada en la parte trasera de la unidad). Asegúrese de que el
interruptor de selección de voltaje (situado encima del interruptor de encendido/apagado) está
ajustado al voltaje correcto del suministro eléctrico. El instrumento debe disponer de una toma de
tierra para garantizar la seguridad eléctrica adecuada. Conecte el cable de alimentación a un
enchufe adecuado según la siguiente tabla.
Conexión
Con corriente
Neutro
Toma de tierra
220/240V, 50/60Hz Supply
marrón
azul
verde/amarillo
110V/120V Supply, 50/60Hz
negro
blanco
verde
3. SÓLO PARA EL REINO UNIDO: El enchufe suministrado con el cable de alimentación incluye un
fusible de 10 amperios para ofrecer protección al instrumento y al usuario.
4. Las unidades cuya placa de régimen indique 230V, 50/60Hz también pueden funcionar entre 210 y
260V; las unidades con 120V, 50/60Hz también funcionan entre 100 y 130V, y las unidades con
100V funcionan entre 90 y 110V. En todos los casos, el porcentaje de calentamiento disminuirá un
8% aproximadamente en los dos extremos del intervalo de voltaje.
5. Conecte el cable de alimentación en el enchufe situado en la parte trasera del instrumento.
6. Sitúe la unidad sobre una mesa plana o en una campana de laboratorio si es necesario, y asegúrese
de que los orificios de ventilación situados en la parte inferior no tienen ninguna obstrucción.
Coloque el instrumento a una distancia mínima de 200mm alrededor de paredes u otros elementos,
y entre cada unidad (en caso de que se utilicen varias) para que haya una circulación de aire
adecuada para cada instrumento.
7. Encienda el instrumento:
I Interruptor de alimentación encendido
O Interruptor de alimentación apagado
CABLE DE REPUESTO
Si es necesario sustituir el cable de alimentación, se debe utilizar un cable de 1mm2 de código
armonizado H05VV, conectado a un enchufe IEC320. EN CASO DE DUDA, PÓNGASE EN
CONTACTO CON UN ELECTRICISTA.
CONDICIONES DE TRABAJO
El ciclador térmico está diseñado para utilizarse en las condiciones siguientes:
•
•
•
•
•
•
Uso en interior
Intervalo de temperatura ambiente +5°C a +40°C
Humedad relativa no superior al 95%
Fluctuaciones del suministro eléctrico no superiores al 10%
Categoría de sobrevoltaje II IEC 60364-4-443
Nivel de contaminación 2
Nota: Las especificaciones de control corresponden a una temperatura ambiental de 20ºC. Las
14
especificaciones pueden empeorar si se utiliza el instrumento fuera del intervalo de temperatura
comprendido entre 10°C y 30°C.
Se han realizado pruebas para comprobar la interferencia de radiofrecuencia del instrumento, el cual
cumple la normativa EN61326.
GARANTÍA
El instrumento está garantizado contra cualquier defecto en el material o la fabricación durante el
período especificado en la tarjeta de garantía que se adjunta. Este período entra en vigor a partir de la
fecha de compra. Durante este período, se reemplazarán sin cargo alguno todas las piezas defectuosas,
a condición que el defecto sea resultado de un accidente, uso incorrecto o negligencia. Tenga en
cuenta que los objetos punzantes, p.ej.: bolígrafos, lápices y uñas, pueden dañar fácilmente la pantalla
del ciclador térmico. Este tipo de daño se considerará como un uso incorrecto, e invalidará la garantía
de este componente.
El distribuidor del instrumento proporcionará información sobre las reparaciones realizadas bajo esta
garantía.
Este manual se ha preparado con una finalidad informativa para los clientes de Bibby Scientific Ltd, y
ninguna parte del manual se deberá considerar como una garantía, condición o reflejo con respecto a
la descripción, comerciabilidad, idoneidad para un fin determinado o de otro tipo de la unidad o sus
componentes.
Con independencia de la descripción y las especificaciones del instrumento que se indican en el manual
del operario, Bibby Scientific Ltd se reserva el derecho de realizar cambios en el instrumento o en
cualquiera de sus componentes cuando lo estime oportuno.
15
INFORMAZIONI SULLA SICUREZZA E L'INSTALLAZIONE
SICUREZZA DELL'OPERATORE
Leggere attentamente il presente manuale prima di usare il ciclatore termico. Se non si usa
l'apparecchiatura nel modo descritto nel presente manuale, la protezione fornita dall'unità potrebbe
risultarne diminuita.
Tutti gli strumenti Bibby Scientific Ltd sono progettati per rispettare i requisiti di sicurezza internazionali
e sono montati con un sezionatore di sovratemperatura. In caso di problemi di sicurezza, spegnere
l'unità dalla presa di rete centrale e togliere la spina dall'alimentazione elettrica.
AVVERTENZA
Le ALTE TEMPERATURE SONO PERICOLOSE in quanto possono provocare serie ustioni agli operatori e
dare fuoco al materiale combustibile. Gli utenti devono conoscere i seguenti pericoli potenziali:
•
•
•
•
PRESTARE ATTENZIONE ED INDOSSARE GUANTI PROTETTIVI PER LE MANI
NON usare sostanze combustibili vicino ad oggetti caldi
NON mettere in funzione lo strumento nei pressi di liquidi o gas infiammabili
NON collocare alcun tipo di liquido direttamente nello strumento, usare sempre le
parti di consumo in plastica idonee
Dopo aver riscaldato i campioni, ricordare che le parti dello strumento, cioè il coperchio ed il blocco ed
eventuali accessori associati potrebbero essere molto caldi.
DISIMBALLAGGIO
Durante il disimballaggio, assicurarsi di aver tolto quanto segue dall'imballo:
•
•
•
•
Ciclatore termico
Cavo di alimentazione di rete
Scheda di garanzia
Certificato di decontaminazione
Si consiglia all'utente di conservare l'imballaggio originale nel caso in cui occorresse restituire lo
strumento per assistenza o riparazioni. Se l'unità non è correttamente imballata e trasportata nel suo
imballo originale, Bibby Scientific Ltd non accetta alcuna responsabilità per eventuali danni che
dovessero verificarsi.
Notare come lo schermo del ciclatore termico si danneggia facilmente con oggetti appuntiti come
penne, matite ed unghia. Questo tipo di danni sarà considerato come uso improprio e renderà nulla la
garanzia per questo componente.
INSTALLAZIONE
Occorre trasportare lo strumento usando entrambe le mani. Non spostare né trasportare lo strumento
quando è in funzione o collegato all'alimentazione elettrica di rete.
1. Tutti gli strumenti Bibby Scientific Ltd sono forniti con un cavo di alimentazione; può essere integrale
o plugin.
16
2. Prima di collegare lo strumento all'alimentazione elettrica di rete, controllare la tensione
confrontandola con la targhetta riportante i valori nominali (si trova sul retro dell'unità). Assicurarsi
che il selettore di tensione (posto sopra l'interruttore On/Off) sia impostato sulla tensione corretta
per l'alimentazione locale. Notare che al fine di garantire la corretta sicurezza elettrica, occorre che
l'unità sia messa a terra. Collegare il cavo di rete ad una presa idonea secondo la tabella riportata
alla pagina successiva.
Connessione
Sotto tensione
Neutro
Terra
220/240V, Alimentazione 50/60Hz
marrone
blu
verde/giallo
Alimentazione 110V/120V, 50/60Hz
nero
bianco
verde
3. SOLO REGNO UNITO: la spina con fusibile fornita con il cavo di rete è dotata di un fusibile da 10
Amp per proteggere lo strumento e l'utente.
4. Le unità la cui targhetta dei valori nominali indica 230V, 50/60Hz funzionano anche tra 210 e 260V;
quelle con 120V, 50/60Hz funzionano anche tra 100 e 130V e quelle contrassegnate da 100V
funzionano tra 90 e 110V. In tutti i casi, il tasso di riscaldamento diminuirà di circa l'8% agli estremi
dell'intervallo di tensione.
5. Inserire il cavo di rete nella presa che si trova sul retro dello strumento.
6. Collocare l'unità su un banco piatto idoneo o in una cappa aspirante se necesario, assicurandosi che
gli sfiati delle prese d'aria nella parte inferiore non siano ostruiti. Posizionare lo strumento con una
distanza minima attorno di 200mm. dalle pareti o da altri oggetti e tra ciascuna unità, nel caso in cui
si usino unità multiple, in modo da consentire un flusso di'aria efficace per ciascuno strumento.
7. Accendere lo strumento:
I Interruttore di rete Acceso
O Interruttore di rete Spento
CAVO DI RICAMBIO
Qualora occorra sostituire il cavo di rete, si dovrà utilizzare un cavo di 1mm2 codice armonizzato
H05VV-F collegato ad una spina IEC 320. IN CASO DI DUBBIO, RIVOLGERSI A UN ELETTRICISTA
QUALIFICATO.
CONDIZIONI DI ESERCIZIO
Il ciclatore termico è stato progettato per funzionare nelle seguenti condizioni:
•
•
•
•
•
•
•
uso interno
range di temperatura ambiente da +5°C a +40°C
altitudine massima 2000 m.
umidità relativa non superiore all'95%
oscillazione dell'alimentazione di rete non superiore al 10%
categoria di sovratensione II IEC 60364-4-443
grado di inquinamento 2
17
Nota: le specifiche di controllo sono indicate ad una temperatura ambiente di 20°C. Le specifiche
potrebbero peggiorare fuori da una temperatura ambiente compresa tra 10°C e 30°C.
Lo strumento è stato collaudato per interferenze da radiofrequenze ed è certificato secondo la norma
EN61326.
GARANZIA
Lo strumento è garantito da qualsiasi difetto nei materiali o nella lavorazione per il periodo specificato
nella scheda di garanzia allegata. Questo periodo è valido dalla data di acquisto; entro tale periodo,
tutte le parti difettose saranno sostituite gratuitamente, a condizione che il difetto non sia la
conseguenza di un incidente, un uso improprio o negligenza. Lo schermo del ciclatore termico può
essere danneggiato da oggetti appuntiti come penne, matite ed unghia. Tale tipo di danno sarà
considerato uso improprio e renderà nulla la garanzia per questo componente.
L'assistenza secondo quanto stabilito dalla presente garanzia deve essere ottenuta dal fornitore dello
strumento.
Il presente manuale è stato preparato ad uso dei clienti di Bibby Scientific Ltd e niente di quanto in esso
contenuto costituisce garanzia, condizione o rappresentanza riguardo la descrizione, la commerciabilità,
l'idoneità allo scopo o altrimenti dell'unità o dei componenti.
Nonostante la descrizione e le specifiche dello strumento contenuti nel manuale dell'operatore, Bibby
Scientific Ltd si riserva il diritto di apportare le modifiche ritenute opportune agli strumenti o a qualsiasi
loro componente.
18
INSTRUMENT CONNECTIONS
Serial Port
RS232/RS485
Selector: Select RS485
for multiple units or
RS232 for a single
unit
Parallel Port
P
RS 485
A
R
A
RS485 Connections
L
RS 485
L
E
RS 485
S
E
R
RS 232
I
A
L
L
D AISY
TERMINATION
CHAIN
BOX
Connection Switch:
Select “Daisy Chain”
for all units except the
last in the chain and
“Termination Box” for
the last unit
Voltage Selector:
Select 230V or 115V
and ensure correct
fuses are fitted
On/Off Switch
Power Lead
Connection: Ensure
that appropriate fuses
are fitted for the
electricity supply
19
CONTACT INFORMATION
For technical, sales or servicing information, contact your local Bibby Scientific Ltd dealer or:
North and South America
Techne Inc.
3 Terri Lane, Suite 10,
Burlington, NJ, 08016, USA
Toll free: 800-225-9243
Tel: +1 609-589-3560
Fax: +1 609-589-2571
e-mail: [email protected]
www.techneusa.com
UK and Rest of World
Bibby Scientific Ltd
Beacon Road, Stone,
Staffordshire, ST15 0SA, UK
Tel: +44 (0)1785 812121
Fax: +44 (0)1785 813748
e-mail: [email protected]
www.bibby-scientific.com
France
Bibby Scientific France SAS
ZI du Rocher Vert - BP 79,
77793 Nemours Cedex,
France
Tel: +33 1 64 45 13 13
Fax: +33 1 64 45 13 00
e-mail: [email protected]
www.bibby-scientific.com
Italy
Bibby Scientific Italia Srl
Via Alcide de Gasperi 56,
20077 Riozzo di Cerro al Lambro,
Milano, Italia
Tel: +39 (0)2 98230679
Fax: +39 (0)2 98230211
e-mail: [email protected]
www.bibby-scientific.it
Middle East
Techne Middle East Ltd.
PO Box 27887, Engomi 2433,
Nicosia,
Cyprus
Tel: +357 22 660427
Fax: +357 22 660356
e-mail: [email protected]
Far East
Bibby Scientific (Asia) Ltd
Room 607, Yen Sheng Centre,
64 Hoi Yuen Road, Kwun Tong,
Kowloon, Hong Kong
Tel: +852 3583 1581
Fax: +852 3583 1580
e-mail [email protected]
www.bibby-scientificasia.com
20
TC-5000 SPECIFICATION
Technical Specification at an ambient temperature of 20°C
Temperature
Gradient Range
Maximum Gradient
Minimum Gradient
Number of temperature probes
Block temperature range
Typical heating rate
Typical cooling rate
Block uniformity (without gradient)
Column uniformity with a 10°C gradient
Control accuracy
Gradient accuracy
Temperature set point adjustment
20°C to 70°C
30°C
1°C
4
4°C to 99°C
3.0°C/sec
1.3°C/sec
<+
_ 0.3°C
<+
_ 0.3°C
<+
_ 0.3°C
+
< _ 0.3°C
+
_ 0.1°C
Heated Lid
The heated lid is only operational if the block temperature is set above 35°C
Selectable heated lid temperature
100°C to 115°C
Heated lid enable/disable
Yes
Warm up time from ambient
4 minutes
Over-temperature cut-out
Yes
Regulated heated lid pressure
Yes
Programs
Password protected
Incremental/decremental hold time
Incremental/decremental temperature
Variable programmable ramp rate steps
Programmable ramp rate
Gradient calculator
Maximum programs stored
Maximum number of segments per loop
Maximum number of loops per program
Program naming capacity
Program interface
Pause facility
End of program alarm
Auto restart on power failure
Run completion time
Yes
Yes
Yes
+
_ 0.1°C
0.1°C
Yes
50
>60
30
Alpha numeric
Touch screen
Yes
Yes (can be disabled)
Yes
Yes
21
Miscellaneous
Height x Width x Length
Weight
Power consumption
Parallel port
Serial port RS232/RS485 adaptable
Connection to PC
260mm x 220mm x 420mm
12.5kg
620W (including the heated lid)
Yes
Yes
Yes
22
TC-5000 OPERATION AND MAINTENANCE
The TC-5000 has many features which increase the flexibility of the instrument and improve the ease of
use. The following features are described in more detail in this chapter:
•
•
•
•
Gradient
Adjustable heated lid
Adjustable screen
PC connections
This chapter also includes useful information relating to the type of consumables which can be used in
the thermal cycler.
GRADIENT
A gradient of temperatures across the block can be useful in optimising the annealing conditions for
primers; a gradient can be set around a temperature in any step of a program. The set temperature is
the temperature required in the middle columns of the block and the gradient is the variation at the
two extremes of the block; the left hand columns being the coolest and the right hand columns the
hottest.
1
2
3
4
5
6
7
8
9
10 11 12
Schematic diagram
of the 96 well block
A
B
C
D
E
F
G
H
lowest
temperature
set temperature
highest
temperature
gradient
The maximum temperature gradient which can be set is 30ºC; the lowest temperature is 20ºC and the
hottest temperature is 70ºC. Examples are given in the table below:
Lowest temperature (ºC) Set temperature (ºC) Highest temperature (ºC)
20
35
50
40
55
70
45
55
65
50
55
60
23
Gradient (ºC)
30
30
20
10
It is important to remember that the greater the gradient, the poorer the uniformity across the block,
therefore fine-tuning may be required to establish the optimum temperature. While the gradient is
mainly linear there are small variations at the extremes of the gradient due to a loss of heat from the
hottest part of the block and a gain of heat from the adjacent column on the coldest. This is the
reason why the first column does not reach the lowest ideal temperature of the gradient and the last
column cannot reach the highest temperature.
A gradient calculator is included in the software which can be used to display more accurately the
temperatures of the block for any given gradient. The gradient calculator is located in the System User
main menu.
ADJUSTABLE HEATED LID
The heated lid of the TC-5000 can be switched on or off depending on the user's requirements. Details
for editing the default lid parameters and those of an individual program can be found on pages 42
and 35 respectively.
The lid is unlocked by lifting the back of the handle as shown in the diagram below, this handle can
then be used to lift the lid. To close, simply lower the lid onto the block containing the samples and
press the handle shut.
TC
-5
0
TC
00
-5
00
0
The pressure of the heated lid must be adjusted for each consumable type used in the thermal cycler,
to ensure that sufficient pressure is present to prevent any evaporation of samples.
Prior to setting the lid pressure, obtain the lowest pressure by rotating the orange disc anticlockwise.
Then shut the lid and latch it. To obtain the correct pressure adjust with the lid in the down position by
gently rotating the orange disc clockwise until it is possible to just feel the pressure being applied.
Finally, rotate the orange disc a further quarter of a turn; the lid is now at the correct pressure for use
with the consumable. The pressure does not need to be adjusted again, unless a different thermal
block or consumable is used.
24
TC
-5
0
00
A heated lid mat (Part No. 6104978) is supplied with the TC-5000; this is for use with 96-well PCR
plates sealed with an adhesive seal. The rubber mat should be placed in top of the sealing film to
ensure good thermal contact with the heated lid. Do not over-tighten the lid as this will cause the
silicone rubber membrane to deform and will reduce the effectiveness of the heated lid.
ADJUSTABLE SCREEN
The brightness of the screen can be adjusted according to the light conditions in the laboratory.
The brightness control is located under the front cover of the instrument. After lifting the front cover
the small black knob in the centre is rotated clockwise to make the screen brighter and anticlockwise
to make it darker.
CONSUMABLES
Bibby Scientific Ltd does not recommend any specific tube or reaction vessel other than those described
in this manual. The tubes must be able to withstand a pressure of 1 atmosphere at 100°C in addition
to the temperatures being used in the thermal cycler block. The tube must not deform to the point of
fracture.
We recommend using reaction volumes between 10µl and 50µl. To test the tube, put 25µl of water in
each of 5 tubes and subject them to a typical program. At the end of the program measure the volume
using a pipette. A loss of more than 1-2µl indicates sample evaporation and a vapour leak. The
volume lost and the change in reagent concentrations determines the minimum volume that can be
used. Typical volume losses of 1µl in 30 cycles allow the use of samples of 10µl or less.
25
PC CONNECTIONS
It is possible to connect either a single thermal cycler or a series of instruments to a computer. The
options are:
1. To connect a single thermal cycler to a computer part number FGEN232 is required which contains a
connecting cable, software disk and instructions.
2. To connect several units via a daisy chain to a single computer part number FTGEN485 is required
which contains an RS485 set. Included are the connector for the computer, 1m cable for connecting
the first instrument to the computer, software disk and instructions. For each unit in the chain a
connecting cable (Part number 6103557) is required.
On each unit select RS485 (up position)
Adapter (if necessary)
Computer
Cables
Computer connector/
RS485 converter
Select “Daisy Chain”
26
Select “Termination Box”
CLEANING THE TC-5000
Prior to cleaning the thermal cycler it is important to disconnect the unit from the power supply and if
it has been in use allow both the block and heated lid to cool to below 50°C prior to attempting to
clean it.
The peltier block, in particular the sample wells should be cleaned regularly to ensure that the
maximum heat transfer from the block to the sample occurs. The block should be cleaned if a spillage
occurs before the instrument is used again. Use a cloth or cotton bud dipped in 50% propan-2-ol
solution (diluted with water) to clean the block ensuring that no deposits are left in the wells of the
block.
The case and screen of the thermal cycler can be cleaned using water, ethanol, hexane or 50%
methanol (diluted with water). Neither the case nor screen should be immersed in the liquid. Solvents
such as acetone should not be used and neither should abrasive cleaners.
If the thermal cycler is being used with any radioactive substances, the peltier block which is made of
aluminum, should be cleaned with an agent which is suitable for nonferrous metals, such as Neutracon
(from Decon Labs Ltd). However, remember that other parts of the unit are made of ferrous materials
and may be damaged by spillage onto them. Neutracon should be used according to the
manufacturer's instructions.
27
REMOVING THE BLOCK
The thermal cycling block can be removed for detailed cleaning or replacement.
First ensure that the unit is disconnected from the power supply and has been allowed to cool if the
unit was previously in use. The two quick release handles located at the front and back of the block are
used to unlock the block. The padlock symbols indicate the direction in which to slide the handles:
open to unlock and closed to lock. When the block is unlocked it will simply lift out of the unit. Never
lift or carry the block by one quick release handle, always use both handles or support the block from
underneath.
When refitting the block, slide the quick release handles in the opposite direction to lock the block into
the thermal cycler. The software will register and calibrate the unit for the new block.
It may be necessary to edit the heated lid parameters for a new application; for example if the in-situ
PCR block is fitted for use with hybridisation slides then it is necessary to switch the heated lid OFF as
excess heat can cause evaporation of samples.
TC
-5
00
0
Removal of the TC-5000 thermal block
28
PROGRAMMING THE TC-5000
START UP MENU
When the TC-5000 is first switched on the screen shown below is displayed and will remain until the
unit has finished performing a series of self tests. This screen displays information such as the serial
number of the instrument, the type of block being used and the version of software running the
thermal cycler.
Techne
TC-5000
Serial number
Software version
Hardware version
Block ID
Last system test
Comms address
Copyright protected 2003
xxxxxx/xx
v.vv
h.hh
FSG - 60x0.5ml
dd/mm/yyyy
01
Performing self tests
0 1 2 3
After the self tests have been completed the
screen opposite is shown unless the unit is
powered up by auto-restart. If there is a
program running and a power failure occurs,
the auto-restart function, if enabled, will
immediately restart the program once power is
restored. The instrument does not perform the
self tests at this point. Once the program is
complete and has been stopped then the self
tests will be initiated.
Techne
TC-5000
Serial number
Software version
Hardware version
Block ID
Last system test
Comms address
Copyright protected 2003
System
User
xxxxxx/xx
v.vv
h.hh
FSG - 60x0.5ml
dd/mm/yyyy
01
System
Admin
The System User section of the software allows
the user to program the thermal cycler for
his/her experiments and view or print the data
logs. The System Admin contains the settings
for the instrument itself.
29
THE SYSTEM USER MENU
In this section of the software the user is able to write new programs and edit existing ones.
Additionally the program defaults for parameters such as the heated lid and final extension steps can
be altered. The gradient calculator and data log files can also be accessed through their sub-menus.
To access the programming software press the
"System User" button on the start-up screen.
The System User menu will then appear.
Techne
TC-5000
Serial number
Software version
Hardware version
Block ID
Last system test
Comms address
Copyright protected 2003
System
User
xxxxxx/xx
v.vv
h.hh
FSG - 60x0.5ml
dd/mm/yyyy
01
The options within the System User menu
include:
System
Admin
Program Defaults: Here the user is able to
edit the default settings for the heated lid,
initial denaturation, final extension, final hold
and the hot start functions, which will then
automatically be inserted into any future
programs.
Programs: The main functions are reading,
writing, editing and running of new or
previously stored programs.
Techne
TC-5000
Gradient Calculator: Calculates the
temperature for each column of the block for
a user-specified temperature and gradient.
Exit: Returns to the start-up menu.
Programs
Program
Defaults
Gradient
Calculator
Exit
30
WRITING AND EDITING PROGRAMS
There are three options available for creating programs on the TC-5000:
1. Copy and edit one of the pre-written templates.
2. Edit a pre-written template and save it using a different name.
3. Create a new program with user-specific parameters.
There are 5 pre-written templates. These templates cannot be altered; however, they can be copied,
given a new name and then edited to save time when writing a new program.
The Ice Bucket and Ligation 15 Degrees programs are simple temperature holds, with the former
having an infinite hold at 4°C and the latter at 15°C. In both of these programs the heated lid is
switched off and the initial denaturation, hot start and final extension functions disabled.
The 2 Step and 3 Step PCR templates along with the RT PCR template have the following default
parameters:
Parameter
2 Step PCR
3 Step PCR
RT PCR
Heated Lid
Heated lid before program
Pause before program
Initial denaturation
Hot start
Final extension
Final hold
105°C
On
Off
05m00s 94.0°C
Disabled
05m00s 72°C
10°C
105°C
On
Off
05m00s 94.0°C
Disabled
05m00s 72°C
10°C
105°C
On
Off
05m00s 94.0°C
Disabled
07m00s 72°C
10°C
The thermal cycling conditions are different for each of these three templates:
2 Step PCR
Cycles
Segment
Segment
Cycles
Segment
Segment
Segment
30
00m30s 92.0°C
01m00s 68.0°C
3 Step PCR
RT PCR
30
00m30s 92.0°C
00m30s 55.0°C
00m30s 72.0°C
1
40m00s
05m00s
40
01m00s
00m50s
01m00s
45.0°C
94.0°C
94.0°C
55.0°C
72.0°C
The RT PCR template is for those experiments where the starting material is RNA rather than DNA.
RNA requires reverse transcription by a reverse transcriptase enzyme into complementary DNA (cDNA)
prior to the amplification by the DNA polymerase in PCR. The extra 45°C step is for the reverse
transcription and the 94°C step is to denature the reverse transcriptase before the PCR.
31
OPTION 1: COPYING AN EXISTING
TEMPLATE
Techne
TC-5000
To access the program templates select
"Programs" in the System Users menu.
Programs
Program
Defaults
Gradient
Calculator
Exit
The cursor arrows can be used to scroll up
and down the stored programs. An individual
program is opened by pressing on the
individual program box (shown opposite).
Select Program
14526 bytes of memory free
b 2 STEP PCR TEMPLATE
b 3 STEP PCR TEMPLATE
Exit
The programs are pre-fixed by b or i. The b
corresponds to the pre-programmed
templates and i for those stored in the
internal memory. Selecting "New" will open
an empty program. See page 35 for more
details.
00:00:00
Once the template is open select "Copy". It is
also possible to run or print the program by
pressing the appropriate button.
b ICE BUCKET
b LITIGATION 15 DEGREES
b RT PCR TEMPLATE
▼
▼
New
Program
3 STEP PCR TEMPLATE
Last Modified
01/11/1998
Heated lid
105.0°C
Heated lid before program
On
Pause before program
Off
Initial denaturation
05m00s
94.0°C
Hot Start
Disabled
Number cycles 30
▼
▼
Copy
Run
Delete
Edit
Print
Exit
It is then necessary to enter a new program
name. There are several text features within the
text screen. Pressing the following buttons
allows access to various options:
Enter the new program name
Q W E R T Y U
I
O P
Num: Numerical keys
Cap: Capital and lowercase letters
BS: Backspace, removes the previous character
Arrow cursors: Move the cursor backwards
and forwards
Space: Introduces a space.
A S D F G H J K L
Cap
Z X C V B N M
➔
Num
Space
➔
Enter
BS Cancel
When finished press "Enter" to return to the
Select Program menu.
32
Select the copied program.
Select Program
14526 bytes of memory free
b ICE BUCKET
b LITIGATION 15 DEGREES
b RT PCR TEMPLATE
i B ACTIN
i GAPDH
▼
▼
New
Program
Exit
The program cannot be changed at this point
so press "Edit" to make any changes
required.
B ACTIN
Last Modified
15/05/2006
Heated lid
11:29:17
105.0°C
Heated lid before program
On
Pause before program
Off
Initial denaturation
05m00s
The program can be copied, deleted, printed
or run by selecting the appropriate option.
94.0°C
Hot Start
Disabled
Number cycles 30
▼
▼
Copy
Delete
Edit
Print
Exit
Run
Editing
All of the parameters can be edited simply by
selecting each line in turn. The cursors can be
used to navigate through the program as all
the parameters cannot be viewed at the same
time. To save any changes select "Save" and a
prompt will ask:
B ACTIN
Initial denaturation
05m00s
Hot Start
94.0°C
Disabled
Number cycles 30
Seg
MAX°C/s
00m30s
92.0°C
Seg
MAX°C/s
00m30s
55.0°C
Seg
MAX°C/s
00m30s
72.0°C
Save
Next step
▼
▼
PWord
Ins
Del
Save As
Save
Yes
No
Exit
and exit back to the program. For details of all
the features and parameters see pages 35.
33
OPTION 2: EDITING AND SAVING AN
EXISTING TEMPLATE
Select Program
14526 bytes of memory free
Select the template from the program menu.
b 2 STEP PCR TEMPLATE
b 3 STEP PCR TEMPLATE
b ICE BUCKET
b LITIGATION 15 DEGREES
b RT PCR TEMPLATE
▼
▼
New
Exit
Select "Edit"
Program
3 STEP PCR TEMPLATE
Last Modified
01/11/1998
00:00:00
Heated lid
105.0°C
Heated lid before program
On
Pause before program
Off
Initial denaturation
05m00s
94.0°C
Hot Start
Disabled
Number cycles 30
▼
▼
Copy
Run
Editing
Delete
Edit
Print
Exit
All of the parameters can be edited simply by
selecting each line in turn. The cursors can be
used to navigate through the program as all
the parameters cannot be viewed at the same
time. Once all the changes have been made,
select "Save As"; there is no option to save.
For details of all the features and parameters
see page 35.
3 STEP PCR TEMPLATE
Initial denaturation
05m00s
94.0°C
Hot Start
Disabled
Number cycles 30
Seg
MAX°C/s
00m30s
92.0°C
Seg
MAX°C/s
00m30s
55.0°C
Seg
MAX°C/s
00m30s
72.0°C
Next step
▼
▼
Ins
PWord
Del
Save As
Exit
Enter a new program name. There are several
text features within the text screen. Pressing
the following buttons allows access to various
options:
Enter the new program name
Q W E R T Y U
I
O P
Num: Numerical keys
Cap: Capital and lowercase letters
BS: Backspace, removes the previous character
Arrow cursors: Move the cursor backwards
and forwards
Space: Introduces a space.
A S D F G H J K L
Cap
Z X C V B N M
➔
Num
Space
➔
Enter
BS Cancel
When finished press "Enter" to return to the
program. Select "Run" to start the program.
34
OPTION 3: CREATING A NEW PROGRAM
Select Program
14526 bytes of memory free
The final option is to create a completely new
program by selecting "New" in the Select
Program menu. This section also contains
details of the individual parameters and
functions.
b 2 STEP PCR TEMPLATE
b 3 STEP PCR TEMPLATE
b ICE BUCKET
b LITIGATION 15 DEGREES
b RT PCR TEMPLATE
To create a new program, select "New" in the
Select Program menu.
▼
▼
New
Editing
Type in a program name; for details see the
previous page.
Exit
The cursor icons can be used to scroll up and
down to access more parameters. Scrolling up
the menu displays the default parameters:
B ACTIN
Initial denaturation
Disabled
Hot Start
Disabled
•
•
•
•
•
•
Next Step
Final extension
Disabled
Final hold
Disabled
▼
▼
PWord
Ins
Del
Save As
Save
Heated lid
Pause before the program
Initial denaturation
Hot start
Final extension
Final hold
The default parameters are defined in the
System user menu and information about the
programming of these parameters can be
found on page 42. All of the default
parameters can be set uniquely within an
individual program.
Exit
The keys in the editing menu are as follows:
"Del"
"Ins"
"Save As"
"Save"
"PWord"
"Exit"
Delete will remove only the highlighted step. The default parameters cannot be deleted.
Insert will add a next step if no "Next step" is present.
Allows the program to be saved with an alternative name.
Saves the program.
Adds a password which protects the program so other users cannot change it.
This returns the screen to the Select Program menu.
The Next step line is inserted automatically
between the Hot Start and Final extension.
Select "Next Step" to open the Edit program
function menu. If Next Step is not displayed
then selecting "Ins" will add a Next Step
below the highlighted line.
Next Step
35
The next step enables one of the following to
be added to the program:
Edit program function
Next step
Segment
7
8
9
➔
Number
Cycles
4
5
6
Continue
Inc
Dec
1
2
3
Cancel
Pause
0
•
BS
OK
Segment: Enables the temperature, time and
heating rate to be inserted into the segment.
Additionally, a gradient across the block can
be added.
➔
Number Cycles: Up to 99 cycles can be
added
Inc Dec: Allows increasing and decreasing
temperatures and times to be set. See page
37 for further details.
Pause: Adds a pause as a step. User
intervention will then be required before the
program can move on to the next step.
INSERTING A SEGMENT
Selecting "Segment" inserts a temperature
step of 94°C for 30s at the maximum ramp
rate.
Edit program function
Segment 94°C
0°C
00m30s
MAX 0°C/s
Segment 94°C
7
8
9
➔
Number
Cycles
4
5
6
Continue
Inc
Dec
1
2
3
Cancel
Pause
0
•
BS
OK
Segment
➔
0°C
00m30s
MAX 0°C/s
If required change the temperature using the
numerical keypad. Then use the ➔ to move
across the screen to the gradient option
0°C.
The gradient on the TC-5000 can be set from 1 to 30°C across the temperature range 20 to 70°C.
Outside this range the accuracy of the different columns cannot be guaranteed. If a gradient greater
than 30°C or outside the temperature range is set then the software will automatically set it to the
maximum, 30°C or within the permitted range. If temperatures are set outside 20 to 70°C range then
the following error message is displayed:
WARNING! Gradient has been
adjusted to be within cycler
limits
Yes
Further information relating to the gradient function can be found on page 23. Use the ➔ to move
across the screen to set the time.
36
The time can be set in hours (h), minutes (m) or
seconds (s). Type in the time required followed
by the unit. Use the ➔ to move across the
screen to the ramp rate.
Edit program function
Segment 94.0°C
0°C
00m30s
MAX 0°C/s
Segment
h
7
8
9
➔
Number
Cycles
m
4
5
6
Continue
Inc
Dec
s
1
2
3
Cancel
0
•
BS
OK
Pause
➔
The ramp rate is automatically set to the
maximum 3°C per second. A number less than
2°C/s can be inserted. Selecting "Max" will
reset the ramp to the maximum.
Edit program function
Segment 95.0°C
0°C
00m15s
MAX 0°C/s
7
8
9
➔
Number
Cycles
4
5
6
Continue
Inc
Dec
1
2
3
Cancel
Pause
0
•
BS
OK
Segment
Max
➔
Selecting "OK" leaves the Edit program
function menu and returns to the program.
Selecting "Continue" adds another new
step to the program.
If a segment or the Inc/Dec function is inserted
then the software will automatically assign 1
cycle to this stage or loop, as it is referred to in
the software.
INSERTING CYCLE NUMBERS
Edit program function
Selecting "Number Cycles" allows the number
of cycles to be increased up to 99. This function
allows the user to easily program the 2 or 3
step section of the PCR which usually involves
between 20 and 45 repeats of 2 or 3
temperature steps. The program will
automatically repeat in the programmed
sequence each of the temperature steps for the
required number of cycles.
Number cycles 01
Segment
7
8
9
➔
Number
Cycles
4
5
6
Continue
Inc
Dec
1
2
3
Cancel
Pause
0
•
BS
OK
➔
INSERTING INCREMENTAL OR DECREMENTAL TIME OR TEMPERATURE
Under normal circumstances, the Hold Temperature of all steps is constant. However, it is possible to
automatically increase or decrease the temperature of a specified step of a programme. Touchdown
amplification of DNA requires a higher temperature during the initial cycles to ensure that only the
specific piece of DNA is amplified and not non-specific products. A decreasing, increasingly less stringent
annealing temperature is then used for the later cycles of the PCR to increase the yield of the product.
37
Temperature
Cycle 1
Cycle 2
Cycle 3
Cycle 4
Decremental temperature for
Touchdown DNA amplification
-i
-i
-i
Time
The Hold Temperature of the incremented/decremented step is the defined Hold Temperature plus the
summation of the increments/decrements. The first cycle is never altered, only subsequent cycles. If
decrement is selected, the Hold Temperature is prevented from falling below 4°C. If increment is
selected the Hold Temperature is prevented from rising above 99°C.
Similarly, under normal circumstances, the Hold Time of all steps is constant. However, it is possible to
automatically increment or decrement the duration of a specified step of a programme. When
amplifying large fragments of DNA it is necessary to ensure that the extension is fully complete after
each cycle, so the extension time is increased with each cycle to compensate for the increased product
quantity and decreasing concentrations of reagents.
Cycle 2
Cycle 3
Temperature
Cycle 1
Incremental time for amplification
of long DNA products
Time
The Hold Time of the incremented/ decremented step is the defined Hold Time plus the summation of
the increments/decrements. The first cycle is never altered, only subsequent cycles. If you select a
decrement, the Hold Time is prevented from falling below one second.
38
INSERTING A PAUSE
Selecting "Pause" adds a stop to the
program, which when running requires user
intervention before the program can
continue.
Edit program function
PAUSE
Segment
7
8
9
➔
Number
Cycles
4
5
6
Continue
Inc
Dec
1
2
3
Cancel
Pause
0
•
BS
OK
➔
SAVING CHANGES TO A PROGRAM
Editing
To change a saved program, select the
program and when opened select "Edit" to
change the parameters.
B ACTIN
Heated lid
105.0°C
Heated lid before program
On
Pause before program
Off
Initial denaturation
10m00s
Hot Start
Select "Save" to save the changes under the
existing program name or "Save As" to save
the changes as a new program.
75.0°C
95.0°C
Next step
Final extention
▼
▼
PWord
07m00s
Ins
Del
Save As
Save
72.0°C
Exit
RUNNING A PROGRAM
To run any previously saved program simply follow these steps:
Select "Programs" from the main System User
menu.
Techne
TC-5000
Programs
Program
Defaults
Gradient
Calculator
Exit
39
Select the program required, using the cursor
keys to view all the available programs.
Select Program
14526 bytes of memory free
b 2 STEP PCR TEMPLATE
b 3 STEP PCR TEMPLATE
b ICE BUCKET
b LITIGATION 15 DEGREES
b RT PCR TEMPLATE
▼
▼
New
Exit
Press "Run".
Program
3 STEP PCR TEMPLATE
Last Modified
01/11/1998
Heated lid
00:00:00
105.0°C
Heated lid before program
On
Pause before program
Initial denaturation
Off
05m00s
94.0°C
Hot Start
Disabled
Number cycles 30
▼
▼
Copy
Run
Delete
Edit
Print
Exit
The run screen is then displayed; if the heated
lid is on then the screen will show the
message "Waiting Heated Lid" and the time
until the lid will be ready. It is possible to stop
or pause the program.
B ACTIN
Program
Waiting Heated Lid 03m40s
If "Stop" is pressed then it is necessary to
confirm that that the program is to be stopped
by pressing "Stop" again.
Stop
If pressing "Stop" was an error then pressing
"Continue" will enable the program to
continue from where it was stopped.
Pause
If "Pause" was selected then press "Unpause"
to restart the program. The pause function
might be required for a manual hot start.
40
As the program continues the temperatures
are displayed on the screen, as shown
opposite.
B ACTIN
Program
Cycle 3 of 3. Step 2 of 3
Holding at 55.0°C
10°C
Sample Lw= 50.0°C, Sample Up= 60°C
Time to complete 01m26s
100
50
0
00h01m
00h02m
Stop
00h03m
00h04m
00h05m
Pause
Program
Program complete
Sample = 72°C
Time to complete 00m00s
This screen is displayed when the run is
complete. If there is a final hold in the
program press "Stop" to end the program.
B ACTIN
100
50
0
00h01m
Stop
Program
Run
00h02m
00h03m
00h04m
00h05m
Pause
Selecting "Run" at this point will start the
program from the beginning again and "Exit"
closes the run screen and displays the
selected program again.
B ACTIN
Exit
41
100
This screen is displayed if there is a power loss
during the running of the program and the
auto-restart function is switched on. The
display shows the times at which the power
failed and resumed.
50
The "Show Info" button takes the screen
back to the program information and display.
Program
Auto Restart
Off 15/05/2006 10:21:28
On 15/05/2006 10:21:36
0
00h01m
00h02m
00h03m
Stop
B ACTIN
00h04m
Pause
When the program has finished or been
stopped then the instrument will perform its
normal start-up tests.
00h05m
Show Info
PROGRAM DEFAULTS
It is possible to change the pre-defined parameters which are used for every program by editing the
Program Defaults. This information can be found in the System User menu.
Techne
TC-5000
Programs
Program
Defaults
Gradient
Calculator
Exit
To access all the parameters it is necessary to
move up and down the menu using the
cursor arrows. The parameters which can be
edited are:
Set program defaults
Heated lid
Disabled
Heated lid before program
Off
Pause before program
Off
Initial denaturation
Disabled
Hot Start
Disabled
▼
▼
Heated lid: Select the "Heated Lid" and then
in the temperature editing window it is
possible to enable/disable the heated lid. If
"Enabled" is pressed the text changes from
xxxxx to allow a temperature to be typed in.
The lid can be programmed between 100 and
115°C.
Exit
Heated lid before program: Selecting this
option simply moves between On and Off.
This option ensures that the heated lid is at
the correct temperature before starting the
actual program.
42
Pause before program: This allows the insertion of a pause in the experiment before the experiment
starts. After the pause there must be user intervention before the program can move on to the next
step of the program.
Initial denaturation: Many experimental
protocols require a high temperature
incubation to remove any secondary structure
of the nucleic acid template.
Edit initial denaturation setting
Initial denaturation
05m00s
94°C
h
7
8
9
➔
m
4
5
6
Enable
s
1
2
3
Cancel
0
•
BS
OK
➔
Selecting "Enable" allows a temperature and
time to be added. The cursors are used to
move between the time and the temperature.
On the left of the numbers there are 3
buttons for setting the time: h for hours, m
for minutes and s for seconds. The correct
button should be pressed after the
temperature has been typed.
Edit initial denaturation setting
Initial denaturation
xxxxxx
xxxxx°C
h
7
8
9
➔
m
4
5
6
Enable
s
1
2
3
Cancel
0
•
BS
OK
➔
Hot start: Many standard enzymes used in molecular biology techniques can hybridise to and extend
non-specific products, resulting in multiple bands on an agarose gel, a greatly reduced yield or no
amplification of the specific product. The 'Hot Start' programming step is used to pause the instrument
at a specific temperature, typically greater than 70ºC, after the initial template denaturation. The
reason is to allow the manual addition of unmodified Taq DNA polymerase which may loose activity if
added during the initial 5 minute denaturation. Heat-activated Taq or Hot Start enzymes do not require
this step.
Final Extension: Many PCR experiments require a final step at the end of the cycling phase to allow
the polymerase to complete the synthesis of the DNA strands. By selecting "Enable" it is possible to
put a time and temperature for the step. A typical example is 72°C for 7 minutes.
Final Hold: Frequently the samples are required for further analysis such as agarose gel electrophoresis
and the samples are better stored at a temperature lower than ambient. Selecting "Enable" allows the
samples to be held infinitely between 4 and 99°C. This step requires the user to stop the program
manually on the instrument. Samples are normally held between 4 and 10°C.
43
GRADIENT CALCULATOR
The gradient is set within the program (see page 36 for further details), however, it is not possible to
view the actual temperatures of each column in the program. The gradient calculator calculates the
temperatures for each of the 12 columns (24 for the 384 well block) and allows the user to determine
the optimal amplification conditions for the assay.
The Gradient Calculator is found in the
System User menu.
Techne
TC-5000
Programs
Program
Defaults
Gradient
Calculator
Exit
Selecting "Temperature" enables the
temperature of the middle of the gradient to
be set. The gradient can be set between 20
and 70°C.
Gradient Calculator
Temperature 55°C
Gradient 10°C
Selecting "Gradient" allows the range of the
gradient to be set. The minimum is 1°C and
the maximum is 30°C.
50.4
51.1
52.2
Temperature
Gradient
53.6
54.5
55.3
Print
56.3
57.6
58.9
An example for the 60 x 0.5ml block is
shown. The gradient is 10°C with the middle
of the block set to 55°C, so the block will be
approximately 50°C on the left and 60°C on
the right and 55°C in the middle of the block.
59.6
Exit
It is possible to print the column and gradient
information by selecting "Print". Selecting
"Exit" returns the screen to the System User
menu.
44
SYSTEM ADMINISTRATION
This section of the software contains settings for the instrument, rather than the actual programs.
From various sub-menus it is possible to change passwords for programs and the instrument, test the
instrument, delete current programs stored on the memory card and completely reset the instrument.
To access the System Admin simply press the
"System Admin" button on the start-up
screen.
Techne
TC-5000
Serial number
Software version
Hardware version
Block ID
Last system test
Comms address
Copyright protected 2003
System
User
xxxxxx/xx
v.vv
h.hh
FSG - 60x0.5ml
dd/mm/yyyy
01
System
Admin
The software will then ask for a password.
Type in the default password TECHNE using
the touch screen as shown opposite.
Enter password
******
Q W E R T Y U
I
To exit the password screen press "Cancel".
O P
Pressing "Enter" takes the user through to
the system administration menu.
A S D F G H J K L
Num
➔
Cap
Z X C V B N M
Space
➔
Enter
BS Cancel
The System Administration Menu.
Techne
TC-5000
Sys Admin
Password
Program
Passwords
Controller
Reset
System
Test
System
Config.
Exit
45
The following functions are located within the System Administration menu:
Program Passwords: Enables an administrator to change the password for any program.
Sys Admin Password: Allows the administrator to change the administrator password.
System Test: Initiates a simple diagnostic test.
Controller Reset: Allows the user to restore the factory settings.
System Configuration: This sub-menu allows the user to change the format of dates, times and
temperature units. Language, auto restart settings and the comms address can also be altered.
Additionally information relating to the heated lid and cycle count of the block is stored in this section
of the software.
PROGRAM PASSWORDS
Select Program
Selecting the "Program Passwords" function
allows the user to change the passwords for
any password protected program. First touch
the screen to select the program for which
the password needs to be changed and then
select "Change PWord". To change the
password simply type the new password by
touching the letters/numbers and then
confirm by pressing "Enter". It is then
necessary to confirm the new password by
re-typing it.
b 2 STEP PCR TEMPLATE
b 3 STEP PCR TEMPLATE
b ICE BUCKET
b LITIGATION 15 DEGREES
b RT PCR TEMPLATE
▼
▼
Change
PWord
Exit
SYSTEM ADMINISTRATION PASSWORD
New system administrator password
Q W E R T Y U
I
Selecting "Sys Admin Password" enables the
user to change the initial administrator
password.
O P
The screen shown opposite is then displayed.
To change the password simply type the new
password by touching the letters/numbers
and then confirm by pressing "Enter". It is
then necessary to confirm the new password
by re-typing it.
A S D F G H J K L
Cap
Z X C V B N M
➔
Num
Space
➔
Enter
BS Cancel
46
SYSTEM TEST
This is a diagnostic test to ensure that the
Real Time Clock settings are correct.
Techne
TC-5000
RTC OK
Exit
CONTROLLER RESET
If "Confirm Reset" is pressed the instrument
default settings will be restored.
Techne
TC-5000
WARNING! All files and programs which
have been stored on the internal memory
will be lost.
*** Controller Reset ***
WARNING!!
This will reset the unit to
system defaults. All stored
files will be lost.
Calibration data is retained
Confirm
Reset
Exit
47
SYSTEM CONFIGURATION
This contains all the settings for programming the instrument such as units of temperature, date
format, language, and date and time settings. Other important features are enabling the auto restart
in case of a power failure and setting the comms address of the instrument if multiple units are linked
to a computer. This part of the software also contains information relating to the cycle count for the
block and the heated lid.
The cursor buttons can be used to scroll up
and down the menu to access additional
parameters.
System Configuration
Temperature units
°C
Beeper
Comms Address
01
Language
English
Date format
dd/mm/yyyy
▼
▼
A parameter is selected by touching the box
on the screen. Then, depending on the
parameter the screen will either toggle
between the options or display the numerical
interface over the page.
On
Exit
To enter the comms address, correct date or
time type in the correct numbers and press
"OK"."BS" (which is short for backspace) and
the cursors can be used to delete and move
across the characters.
Todays Date
Enter date 05/01/2006
7
8
9
4
5
6
1
2
3
Cancel
0
•
BS
OK
➔
➔
The parameters which are editable in the systems configuration menu are:
Temperature units: This allows the user to change the units used in the thermal cycling programming of
the TC-5000. There are 2 options: degrees Centigrade (°C) or degrees Fahrenheit (°F).
Beeper: If required the instrument can be programmed to beep at the end of the program and also when
buttons are pressed on the touch screen. This can enabled or disabled.
Comms Address: This enables the user to specify a unique identification number for each instrument if
multiple instruments are connected together and controlled via a computer. It is possible to select a
communications address between 01 and 64.
Language: There are 5 languages for the user to select from: English, Français, Espanôl, Deutsch and
Italiano.
Date Format: This allows the user to change the date format from dd/mm/yyyy to mm/dd/yyyy.
48
Date: The date can be corrected by the user by using the numerical interface shown above.
Time: The time can be corrected by the user by using the numerical interface shown in above.
Auto restart: This allows a program to be re-started in the event of a power failure when the instrument
is in the middle of running a program. The options are Enabled or Disabled.
Baud Rate: This is the speed of communication between the thermal cycler and a computer. The rate can
be selected from the range of 300 to 38400 depending on individual requirements.
49
FREQUENTLY ASKED QUESTIONS
Q1
What is "Hot start"?
A1
The Hot Start step is used to pause the machine at a specific temperature, typically around 70°C,
after the initial template denaturation. The reason is to allow the manual addition of unmodified
Taq DNA polymerase which may loose activity if added during the initial 5min denaturation.
Heat-activated Taq or Hot Start enzymes do not require this step.
Q2
What is the incremental function? (See page 37).
A2
Incremental timing and temperature are used to increase or decrease either the time or
temperature incrementally over the number of cycles in a stage.
Incrementation of extension time is used with 'Long PCR' which is when large template
fragments are to be amplified (e.g. 27kb lambda DNA, 40kb genomic DNA).
Decremental temperature is used for protocols such as 'Touchdown PCR' where the first cycle
starts with a high annealing temperature and over the number of cycles there is a gradual
decrease in the temperature. This ensures that only the specific product is amplified.
Q3
What is a gradient thermal cycler?
A3
Gradient blocks enable a particular temperature step in a protocol to be altered so that the
temperature varies across the block. By specifying a temperature and the gradient to be applied,
each column will achieve a different temperature, with the set temperature in the middle block,
the lowest on the left and the highest on the right. The gradient is the total difference across the
block, for example if the set temperature was 60°C and the gradient 10°C then the
temperatures would range from 55°C to 65°C from left to right.
Q4
Why is a temperature gradient required?
A4
The annealing temperature of the specific primers used in DNA amplification often requires
optimisation. Instead of running multiple experiments with different annealing temperatures the
gradient thermal cycler allows the testing of 12 different annealing temperatures simultaneously.
If you require further technical or applicational assistance please contact Bibby Scientific Ltd at:
[email protected]
For servicing information please contact: [email protected]
We are continually striving to improve our thermal cyclers and software. If you have any comments and
suggestions on how we can do things better please send them to us at:
[email protected]
50
ADDITIONAL INFORMATION
Brief fault finding notes and a list of replacement parts are given in this section.
Note: that this equipment should only be dismantled by properly trained personnel. Removing the
outer cover exposes potentially lethal mains voltages. There are no user serviceable parts within this
equipment.
FAULT FINDING
Should you have any problems with your TC-5000 which cannot be easily remedied, you should contact
your supplier and return the unit if necessary. Please include details of the fault observed and remember
to return the unit in its original packing. Bibby Scientific Ltd accept no responsibility for damage to units
which are not properly packed for shipping: if in doubt, contact your supplier, giving the full serial
number of the unit and software version number (shown when the unit is first switched on).
FUSES
If neither the power light nor display on the front panel is lit, one of the two fuses may have blown.
Check that there is no external cause, such as a faulty plug or lead. Check both fuses and replace the
faulty fuse with a new one of the correct value (fuse values are given on the label next to the power
inlet). Note that fuses should only be replaced by a qualified electrician.
The holder for the two fuses is built into the mains input socket. First remove the power cable and then
gently prise the fuse drawer open with a flat-bladed screwdriver or similar tool.
Each fuse can be removed by using the screwdriver as a lever.
Exchange the faulty fuse in the fuse holder for a working fuse of the correct value. Finally, replace the
fuse drawer in the fuse compartment and push the drawer shut.
Fuses which blow repeatedly indicate a serious fault and you should return the unit to your supplier for
repair.
ACCESSORIES
Product no
Description
Quantity
FTUB05TW
0.5ml thin wall microtubes
Pack of 1000 tubes with caps
FTUB02TW
0.2ml thin wall microtubes
Pack of 1000 tubes with caps
FMITUBC
0.2ml thin wall microtubes
250 strips of 8 tubes
FMICAPC
0.2ml microtube caps
250 strips of 8 caps
F96T02
96-well flat top, non-skirted plates
Pack of 25 plates
F96PLHS
96-well half skirted plates
Pack of 25 plates
F96SEAL
96-well sealing mat (silicon)
Pack of 50 mats
FMIP384
384-well skirted plates
Pack of 50 plates
FHSEAL
Heat sealing film
Pack of 100
51
Product Number
Description
Quantity
FHSEALSD
Heat sealer
1
FHSPA96
Heat sealer plate adapter for 96-well plates
1
FHSPA384
Heat sealer plate adapter for 384-well plates
1
FTC51BHD
TC-5000 block for 96 x 0.2ml microtubes
or 96-well plate
1
TC-5000 block for 96 x 0.2ml microtubes
or 96-well fully skirted plate
1
FTC51B5D
TC-5000 block 60 x 0.5ml microtubes
1
FTC51BID
Flat plate block for in-situ (non gradient)
1
FTCFB384
TC-5000 block for 384-well fully skirted plate
1
FTC51BFD
PC ACCESSORIES
Product Number
Description
FGEN232
Cable and software to connect 1 cycler
to PC (RS232 only)
1
Power pack, 230V, 485 converter, cable and
software to connect 2+ cyclers to PC
1
FGENONE
One cycler cable extension set
1
FGENTWO
Two cycler cable set
1
FGENFOUR
Four cycler cable set
1
FGENTEN
Ten cycler cable set
1
FGEN485D
Quantity
REPLACEMENT PARTS
Product Number
Description
Quantity Required
FCABRTUK
Mains cable and plug UK 230V
1
FCABRTEU
Mains cable and Schuko plug 230V
1
FCABRTUS
Mains cable and US 3-pin plug 120V
1
FCABRTUS
Mains cable and US 3-pin plug 100V
1
6500342
Fuse T6.3A, 230V units
2
6008445
Fuse T8A, 120V units
2
6500324
Fuse T10A, 100V units
2
52
Bibby Scientific France SAS
Bibby Scientific Italia Srl
ZI du Rocher Vert - BP 79,
77793 Nemours Cedex,
France
Tel: +33 1 64 45 13 13
Fax: +33 1 64 45 13 00
e-mail: [email protected]
www.bibby-scientific.com
Via Alcide de Gasperi 56,
20077 Riozzo di Cerro al Lambro,
Milano, Italia
Tel: +39 (0)2 98230679
Fax: +39 (0)2 98230211
e-mail: [email protected]
www.bibby-scientific.it
Bibby Scientific Ltd
Techne Inc.
Bibby Scientific (Asia) Ltd
Beacon Road, Stone,
Staffordshire, ST15 0SA
United Kingdom
Tel: +44 (0)1785 812121
Fax: +44 (0)1785 813748
email: [email protected]
www.bibby-scientific.com
3 Terri Lane, Suite 10,
Burlington, NJ, 08016, USA
Toll free: 800-225-9243
Tel: +1 609-589-3560
Fax: +1 609-589-2571
email: [email protected]
www.techneusa.com
Room 607, Yen Sheng Centre,
64 Hoi Yuen Road, Kwun Tong,
Kowloon, Hong Kong
Tel: +852 3583 1581
Fax: +852 3583 1580
e-mail [email protected]
www.bibby-scientificasia.com
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