TGGE and TGGE Maxi Family

TGGE and TGGE Maxi Family
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TGGE and TGGE MAXI
Introduction
• Peltier powered linear temperature gradient
• Microprocessor control for reliable assay conditions
• No casting of chemical gradient gels
With the Biometra TGGE system
biomolecules are separated in a
temperature gradient according to their
melting behavior. Unlike chemical
gradients the Peltier driven temperature
gradient can be controlled by a
microprocessor thus providing unmatched
reproducibility. With TGGE PCR fragments
differing in a single position only can be
separated quickly and cost efficiently.
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Precise
The heart of both TGGE systems is the
temperature block which is powered
by Peltier technology. Thanks to precise
microprocessor control a strictly linear
gradient is generated providing a
maximum of reproducibility. Thus assay
conditions can be much better (and
easier) controlled compared to
conventional chemical gradients or
temporal gradients using water baths.
Sensitive
In contrast to direct sequencing TGGE
also detects mutations in mixed DNA
samples. Whenever heterozygous DNA
is to be analysed, direct sequencing will
not give a clear signal for the position of
the mutation. This is especially problematic
when the mutated gene is masked by a
high background of normal cells. TGGE
reliably detects mutations in a 1:10
dilution (and higher) of wildtype DNA.
Cost saving
TGGE provides fast and cost effective
mutation screening of PCR fragments.
Only those samples are selected for
direct sequencing that are different from
the wildtype (standard). The total number
of sequenced samples can be dramatically
reduced saving time and money. Ready
made gels for routine application are
available.
Patented technology
TGGE is protected by patents in most
countries of the world. The patent for
TGGE method and TGGE instrumentation
is held by Qiagen AG, Hilden. Biometra
is the exclusive licensee for manufacturing
and distribution of TGGE instrumentation.
Elect rophoresis TGGE
TGGE and TGGE Maxi System
Temperature Gradient Gel Electrophoresis
Biometra TGGE System
• Rapid separation of mixed
DNA samples
• Quick optimisation of new protocols
The TGGE system was designed for
maximum resolution in a small gel.
The highly integrated system consists of
the gradient electrophoresis unit and an
external controller. Both, optimisation of
temperature range and parallel analysis
of multiple samples can be achieved in
very short time. New protocols a rapidly
established and high serial throughput
can be achieved.
Biometra TGGE MAXI System
• Large format for separation of
complex sample mixtures
• High parallel sample throughput
The TGGE MAXI system provides a
large separation distance for analysis
of complex samples. In addition, a high
number of samples can be analysed in
parallel.
Technical Specifications
see page 68
Ordering Information
see page 69
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Principle of TGGE
During migration through a temperature
gradient DNA starts to melt. Once the
DNA is partially single stranded, the
sample is slowed down dramatically.
Other samples that are still completely
double stranded at this temperature
further migrate through the gel. The
earlier (lower) a DNA melts, the earlier
the sample stops in the gel. Since the
melting temperature of DNA is determined
by the primary sequence, there is a
direct relation between sequence and
melting temperature.
TGGE is ideally suited to separate DNA
fragments of identical size on the basis
of their DNA sequence. This is typically
used for screening of PCR fragments
(of identical length) for mutations.
cold
partial ss
warm
Perpendicular and parallel TGGE: principle of analysis
Perpendicular TGGE (A): One sample is separated over a broad temperature
range (gradient perpendicular to migration of samples). This mode is applied
to identify the optimum temperature range for separation of this sample.
Parallele TGGE (B): Multiple samples are analysed in parallel (gradient
parallel to migration of samples). This mode is applied for routine analysis
of many samples.
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Elect rophoresis TGGE
TGGE in Mutation Analysis
Perpendicular TGGE for optimisation of
temperature gradient
A 120 bp DNA fragment was separated
in a perpendicular temperature gradient.
Prior to analysis the sample has been
mixed with non-mutated standard DNA.
Mixture was denatured at 94 °C and
then slowly cooled down to allow for
heteroduplex formation.
By comparing the melting curve with
the temperature lanes on the block, the
optimum gradient for parallel analysis
of multiple samples is obtained.
Temperature gradient 30 °C (left) to 70 °C (right),
250 V, 45 min.
Parallel analyis of heteroduplex
samples
In contrast to conventional electrophoresis
techniques, in TGGE separation is not
improved by extended running times but
by adjusting the temperature gradient.
In this example a gradient of only 5 °C
was applied to efficiently separate
heteroduplex DNA samples. All bands in
this gel are of identical length (120 bp),
there is only a difference in one position
of the DNA sequence.
Temperature gradient 39 °C (top) to 44 °C (bottom),
150 V, 50 min.
Parallel analysis of many samples with
the TGGE MAXI System
In this example the bands have migrated
to the middle of the gel resulting in a
rather long separation time. To reduce
running time, temperature at the top of
the gel can be elevated. Thus samples
melt already in the top part of the gel
and can be distinguished.
Clean gels for the TGGE MAXI System
(024 - 240) cover only half of the
thermoblock. By using only part of the
separation distance short analysis times
are achieved.
Temperature gradient 35 °C (top) to 60 °C (bottom),
350 V, 4 hours
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TGGE
Technical Specifications
TGGE System
TGGE MAXI System
Peltier
Peltier
5 - 80 °C
5 - 80 °C
Electrophoresis Unit
Technology
Temperature range
Linear gradient
Themoblock dimensions
Glass plate dimensions
Gel dimensions
Separation distance
(perpendicular/parallel)
Sample numbers (volume)
45 °C
45 °C
9 cm x 9 cm
20 cm x 20 cm
9 cm x 9 cm
23.5 cm x 23.5 cm
7.8 cm x 4.2 cm
20 cm x 21.7 cm
4 cm/5 cm
16 cm/19 cm
10 x (5 μl)
32 x (5 μl)
12 x (3 μl)
42 x (5 μl), clean gels (024 - 240)
18 x (1.5 μl)
13 x (5 μl), clean gels (024 - 040)
23 cm x 23 cm x 23 cm
43 cm x 43 cm x 33 cm
Weight
4.2 kg
22.0 kg
Controller
External
External
Control of electrophoresis
Control of electrophoresis
and temperature gradient
and temperature gradient
31 cm x 22 cm x 12 cm
31 cm x 22 cm x 12 cm
3.8 kg
3.5 kg
Power Supply
Integrated in controller
External
Max. Voltage
400 V
400 V
500 mA
500 mA
30 W
50 W
Apparatus dimensions (L x W x H)
Dimensions (L x W x H)
Weight
Max. Amperage
Max. Power
Control modus
Fixed voltage
Weight
68
Fixed voltage
30 cm x 22 cm x 8 cm
Dimensions (L x W x H)
-
6.5 kg
Elect rophoresis TGGE
TGGE
Order Information
Model
Order No.
TGGE System, 230/115 V
024 - 000
Electrophoresis unit with high precision gradient block, 2 buffer chambers for variable positioning,
controller with integrated power supply, starter kit (024 - 003)
TGGE Starter Kit for 25 gels
024 - 003
inlcuding glass plate 1 slot (024 - 023), glass plate 8 slots (024 - 022), glass plate 12 slots (024 - 025),
3 cover plates (024 - 021), 100 buffer wicks (024 - 015), 25 Polybond films, sample AcrylGlide, 3 plastic clamps
TGGE glass plate, 8 slots (5 μl)
024 - 022
TGGE glass plate, 1 slot (50 μl)
024 - 023
TGGE glass plate, 1 diagonal slot (75 μl)
024 - 024
TGGE glass plate, 12 slots (3 μl)
024 - 025
TGGE glass plate, 18 slots (1 – 2 μl)
024 - 026
TGGE glass plate, with 0.5 mm spacer, no slots
024 - 027
TGGE bonding plate, without spacers
024 - 021
TGGE buffer wicks, 7 cm x 7 cm, 100/pkg
024 - 015
TGGE PolyBond film, 8.8 cm x 8.8 cm, 25/pkg
024 - 030
TGGE PolyBond film, 8.8 cm x 8.8 cm, 100/pkg
024 - 034
TGGE cover plate + 10 hydrophobic cover films
024 - 031
TGGE hydrophobic cover film, 7 cm x 6 cm, 25/pkg
024 - 032
TGGE hydrophobic cover films, 100/pkg
024 - 035
TGGE self-adhesive slotformers (10 x multi-well, 9 x long-well)
024 - 121
Testkit for TGGE and TGGE Maxi including 40 μl wildtype DNA, 40 μl mutant DNA, 400 μl heteroduplex DNA,
024 - 050
1ml loading buffer
TGGE casting stand for 5 gels
024 - 028
TGGE clean gels, 8.3 cm x 8.8 cm, 13 slots (5 μl), 10 gels
024 - 040
TGGE MAXI System, 230/115 V
024 - 200
Electrophoresis unit with high precision gradient block, 2 buffer chambers for variable positioning,
controller, power supply, manual, MAXI starter kit (024 - 204)
TGGE MAXI Starter Kit for 25 gels, including glass plate 1 slot (024 - 228), glass plate 32 slots (024 - 229),
024 - 204
2 cover plates (024 - 227), 2 sealings (024 - 230), 25 cover films (024 - 232), 25 Polybond films (024 - 234),
4 buffer wicks (024 - 216), sample AcrylGlide, 12 binder clamps
TGGE MAXI glass plate, without spacer 23.5 cm x 23.5 cm
024 - 221
TGGE MAXI glass plate, with spacer, no slot former
024 - 227
TGGE MAXI glass plate perpendicular, spacer (1 mm) and 1 slot (75 μl)
024 - 228
TGGE MAXI glass plate parallel, spacer (1 mm) and 32 slots (5 μl)
024 - 229
TGGE MAXI silicone sealing for gel casting, 1 mm
024 - 230
TGGE MAXI gel cover film, 25/pkg
024 - 232
TGGE MAXI Polybond film, 25/pkg
024 - 234
TGGE MAXI Polybond film, 100/pkg
024 - 235
TGGE MAXI self-adhesive slotforming units for parallel gels (8 strips with 28 x 5 μl each and 9 x 200 μl)
024 - 222
TGGE MAXI buffer wicks re-usable, 4 pcs.
024 - 216
TGGE MAXI clean gels, 23.3 cm x 11.5 cm, 42 slots (5 μl), 5 gels
024 - 240
TGGE MAXI Applicator strip, silicone, 34 slots for 8 μl (for use with 024 - 227)
024 - 223
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