IFU026_PD5.2-5 SBT Resolver IFU (RUO)

IFU026_PD5.2-5 SBT Resolver IFU (RUO)
PCR Amplification and Sequencing of HLA DRB1 Full Exon 2
Instructions for Use
Version No: 1.0
Issue Date: August 2012
For Research Use Only. Not for use in diagnostic procedures. No claim or representation is
intended to provide information for the diagnosis, prevention, or treatment of a disease.
Conexio Genomics Pty Ltd
8/31 Pakenham St
Fremantle 6160
Western Australia
Australia
Page 1 of 16
For Research Use Only
Contents
PRINCIPLE ............................................................................................................................................ 3
KIT COMPOSITION ............................................................................................................................ 4
STORAGE REQUIREMENTS ............................................................................................................. 5
MATERIALS, REAGENTS AND EQUIPMENT NOT SUPPLIED ................................................. 5
SAMPLE REQUIREMENTS ................................................................................................................ 6
WARNINGS AND SAFETY PRECAUTIONS ................................................................................... 6
PROCEDURE ......................................................................................................................................... 7
1.
2.
3.
4.
5.
6.
7.
PCR ............................................................................................................................................ 7
AGAROSE GEL ELECTROPHORESIS .............................................................................................. 8
PURIFICATION OF PCR PRODUCT ................................................................................................ 8
SEQUENCING REACTION ............................................................................................................. 9
PURIFICATION OF SEQUENCING REACTION PRODUCTS.............................................................. 10
DENATURATION & ELECTROPHORESIS OF SEQUENCING REACTION PRODUCTS ........................ 11
EDITING AND ANALYSIS OF ELECTROPHEROGRAMS .................................................................. 11
PERFORMANCE CHARACTERISTICS ......................................................................................... 12
LIMITATIONS AND CAUTIONS ..................................................................................................... 12
LICENSE .............................................................................................................................................. 12
TROUBLESHOOTING ....................................................................................................................... 13
RELATED PRODUCTS ...................................................................................................................... 15
SUPPORT AND CONTACT DETAILS............................................................................................. 16
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For Research Use Only
Principle
The HLA Sequence Based Typing (SBT) procedure described here, involves the initial
amplification of the target sequences followed by treatment with ExoSAP-IT® to remove
unincorporated primers and dNTPs. The amplicon is then used as a template for direct
automated fluorescent DNA sequencing using customized sequencing primers and the Big
Dye® Terminator sequencing chemistry available from Applied Biosystems™ by Life
Technologies™. The extension products are purified according to the ethanol precipitation
method and denatured using Hi-Di™ formamide available from Applied Biosystems™ by
Life Technologies™, before separation and detection on an automated fluorescent DNA
sequencer. It is recommended that the resulting data is then analysed with Assign™ sequence
analysis software from Conexio Genomics Pty Ltd.
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For Research Use Only
Kit Composition
PRE-PCR Contents†
(No of vials)
Kit
HLA-DRB1
HH-PD5.2-5(20)
HH-PD5.2-5(50)
20 tests
50 tests
DNA POL – DRB1
1 x 9L
DRB1EX2F
HLA-DRB1 MIX
1 x 370L
DRB1EX3R-2
DNA POL – DRB1
1 x 18L
DRB1EX2F
HLA-DRB1 MIX
1 x 920L
DRB1EX3R-2
†
The PRE-PCR kit contains a vial of a locus-specific PCR mix (i.e. HLA-DRB1 MIX
primers, along with a single vial of DNA polymerase (
The POST-PCR kit contains sequencing primers (e.g.
POST-PCR Contents
(No of vials)
DNA POL – DRB1
DRB1EX2F
DRB1EX2R-2
1 x 44L each
RB-TG344-R
DRB1EX2R-2
1 x 110L each
RB-TG344-R
) consisting of PCR buffer, dNTPs, MgCl2, and locus specific PCR
).
).
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For Research Use Only
Storage Requirements
The PRE- and POST-PCR boxes may be separated and stored in designated PRE- and POSTPCR freezers. When stored at -20C (temperature range of -15C to -25C is acceptable), the
kit components can be used until the expiry date indicated on the outer labels and can tolerate
up to 25 freeze-thaw cycles.
To maintain optimal kit performance, the kit components should be removed from the -20C
storage location and thawed rapidly at room temperature before use. The kit components, with
the exception of the polymerase, should then be gently vortexed to ensure that the
components of each tube are appropriately mixed after thawing. After use, the
kits/components should be returned immediately to -20C.
Materials, Reagents and Equipment Not Supplied
PCR
1.
Sterile water
2.
Electronic or mechanical pipettes and aerosol-resistant tips
3.
Thermal cycler with heated lid
These kits have been validated using the following thermal cyclers:
MJ Research PTC 225 DNA Engine DYAD™, Applied Biosystems™ by Life
Technologies™ Gene Amp® PCR System 9700, and Eppendorf Mastercycler® Pro.
Use of other thermal cyclers with these kits requires validation by the user.
4.
0.2mL thin-walled thermal cycling reaction tubes (8 well strips or 96 well plates).
Use those recommended for use with your thermal cycler.
5.
Sterile 1.5mL tubes
6.
Sterile work area such as biological safety cabinet or hood.
7.
Table top centrifuge with plate adapters and capacity to reach 2500 x g
8.
Vortex
Agarose Gel Electrophoresis
9.
Agarose gel electrophoresis apparatus
10.
1% agarose (molecular biology grade) TBE gel containing 0.1g/mL ethidium
bromide.
11.
Loading buffer
12.
PCR Marker suitable to cover range of 300 – 1300 bp
13.
UV transilluminator
Purification of PCR Product
14.
ExoSAP-IT® (USB® Products Cat No 78200 for 100 reactions)
15.
2mM MgCl2 (available for purchase from Conexio Genomics, product code MgCl21.0(50) or MgCl2-1.0(3000))
16.
Shaker
The use of alternative PCR purification techniques requires validation by the
user prior to use.
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For Research Use Only
Sequencing Reaction
17.
BigDye® Terminator Cycle Sequencing Kit v3.1 or v1.1, Applied Biosystems™ by
Life Technologies™.
18.
5x Sequencing Reaction Buffer (Conexio Genomics, product code SEQ BUF2.0(400) or SEQ BUF-2.0(5000)) or BigDye® Terminator v3.1 or v1.1 5X
Sequencing Buffer, Applied Biosystems™ by Life Technologies™.
Purification of Sequencing Reaction Products
19.
125mM EDTA, pH8.0 (available for purchase from Conexio Genomics, product code
EDTA-3.0(200) or EDTA-3.0(5000))
20.
Absolute and 80% Ethanol. Each run requires freshly prepared 80% ethanol
consisting of absolute ethanol and sterile water. DO NOT USE DENATURED
ETHANOL.
The use of alternative sequencing purification techniques requires validation by
the user prior to use.
Denaturation and Electrophoresis of Sequencing Reaction Products
21.
Hi-Di™ Formamide, Applied Biosystems™ by Life Technologies™, product code
4311320
22.
Automated DNA Sequencer and accessories (e.g. Applied Biosystems™ by Life
Technologies™ ABI Prism® 3730), including data collection and software.
These kits have been tested and validated on the Applied Biosystems™ by Life
Technologies™ 3100, 3730 and 3730xl capillary sequencers and software.
The use of other denaturation techniques and sequencing platforms requires
validation by the user prior to use.
23.
HLA Sequencing Analysis Software (e.g. Assign™ SBT, version 3.5 or higher or
Assign™ ATF, Conexio Genomics Pty Ltd).
Sample Requirements
1. Sterile water (negative/ no template control)
2. High molecular weight human genomic DNA (concentration range of 20-100ng/µL in
Tris/EDTA buffer and OD260/280> 1.8) extracted from ACD or EDTA anticoagulated
whole blood specimens. Do NOT use whole blood specimens containing heparin.
Warnings and Safety Precautions

This kit must be used by trained and authorized laboratory personnel.

All samples, equipment and reagents must be handled in accordance with good
laboratory practice. In particular, all patient material should be considered as
potentially infectious. The use of gloves and laboratory coats is strongly
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For Research Use Only
recommended. Handle and dispose of all sample material according to local and
national regulatory guidelines.

There are NO dangerous substances contained in any of the kit components.

Do NOT use reagents beyond their expiration date.

The use of kit components from different kit batches is NOT recommended. Such use
may affect the assay’s performance.

Use of reagents not included in this kit or not listed under “Materials, Reagents and
Equipment Not Supplied” (e.g. alternative Taq DNA polymerases) is NOT
recommended. Such use may affect the performance of the assay.

Care should be taken to prevent cross-contamination of DNA specimens. Change tips
between DNA specimens wherever possible.

Pre- and Post-PCR activities must be strictly physically separated. Use specifically
designated equipment, reagents and laboratory coats.

Ethidium bromide is a potential carcinogen. Protective gloves must always be used
when preparing and handling gels. Dispose of ethidium-bromide gels and buffers
according to local and national guidelines.

While viewing and photographing agarose gels under UV light, always avoid direct
exposure and use appropriate UV-blocking face protection, disposable gloves and
laboratory coats.
Procedure
1. PCR
1.1. A separate PCR reaction will need to be set up for each sample to be tested. Each run
should include appropriate positive control/s of known genotype, and at least one
negative control.
1.2. Prepare a fresh solution of PCR master mix each time a PCR is performed. Quickly
thaw the locus-specific PCR mix at room temperature. Once thawed, vortex briefly.
1.3. Dispense the required volume of PCR mix and DNA polymerase into a sterile tube
for the number of samples to be tested (refer to Table 1 below for the volume per
reaction). Pulse vortex the solution 3-4 times.
Component
Volume
HLA-DRB1 MIX
16.7L
DNA POL – DRB1
0.3L
Table 1: Composition of the master mix required per sample.
1.4. Dispense 17L of the master mix into each reaction well.
1.5. Add 3L of sample DNA or appropriate positive control/s to each reaction well. Add
3L of sterile water to the negative control reaction well.
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For Research Use Only
1.6. Seal the reaction wells. Mix gently by vortexing and centrifuge briefly.
1.7. Place the reaction wells into a thermal cycler and run according to the thermal cycling
conditions below.
95°C - 10 mins
96°C - 20 secs
60°C - 30 secs
72°C - 3 mins
33 cycles
15°C - hold
1.8. Amplification takes approximately 2.5 hours to complete.
1.9. When the PCR is completed, remove the reaction wells/plate from the thermal cycler
and either proceed directly to gel electrophoresis or store at 4°C until required.
NOTE: Purification of amplicons by ExoSap-IT® treatment should occur within 24 hours of
completion of PCR.
2. Agarose Gel Electrophoresis
2.1. Confirm successful amplification by agarose gel electrophoresis using 2L of each
PCR product combined with 5L loading buffer (alternative volumes of loading
buffer should be validated prior to use). The use of 1% agarose gels is
recommended.
The number and expected sizes of the resultant amplicons will vary according to the sample
genotype. Expected PCR amplicon sizes range between ≈ 450 bp - 850bp.
3. Purification of PCR Product
NOTE: Purification systems other than EXOSAP-IT® (e.g. Agencourt® AMPure® XP or
column-based systems) can be used to purify these PCR products. It is strongly recommended
that users validate these procedures before proceeding. If EXOSAP-IT® is to be used it is
recommended that users follow the procedure described below.
3.1. Prepare a mastermix consisting of 4L of ExoSAP-IT® and 8L of 2mM MgCl2 per
sample to be purified. Dispense 12L of the mastermix into the reaction well of each
reactive sample. Seal the wells and vortex to mix. Centrifuge briefly before placing
into the thermal cycler. Run the thermal cycler according to the following profile:
37°C - 30mins
80°C - 15mins
4°C - hold
3.2. Upon completion, dilute the purified product 1:4 with sterile water. This dilution step
will ensure that there is sufficient template to perform the sequencing reactions and
ensure that the concentration of the template is sufficient to produce good quality
sequence data.
NOTE: A higher dilution factor (e.g. 1:8) may be required if consistently high signals and
associated noise and artefacts are observed. Weaker PCR products may require a lower
dilution factor.
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For Research Use Only
3.3. ExoSAP-IT® treated samples may be stored at 4C until ready for use. ExoSAP-IT®
treated samples can be stored at 4C for up to a week before use, but should be
stored at -20C for long term storage.
4. Sequencing Reaction
NOTE: In instances where heterozygous ambiguities are to be resolved with hemizygous
sequencing primers such as HARPS®, please refer to the SBT Resolver™ HARPS®
Instructions for Use.
4.1. Table 2 lists the sequencing primers that are to be used for this kit.
HLA-DRB1†
DRB1EX2F
DRB1EX2R-2
DRB1EX3R-2
RB-TG344-R†
Table 2: Sequencing primers provided for use with the HH-PD5.2-5 kits.
†
®
RB-TG344-R is a HARP directed to the codon 86 dimorphism. Its use is optional.
4.2. Prepare a fresh solution of sequencing primer mix on ice each time a sequence
reaction is performed. The composition and volumes for the mix indicated below are
per sample.
Component
Sequencing primer
Volume
2 µL
Sterile water
11.5 µL
®
BigDye Terminators
1 µL
5x Seq Rxn Buffer
3.5 µL
4.3. Mix each sequencing reaction mixture gently by pulse vortexing.
4.4. Dispense 18µL of the sequencing reaction mix into each appropriate reaction well.
NOTE: For runs which involve few samples with many sequencing primers, it is
acceptable to dispense the sequencing primer (2µL) directly into the individual
reaction wells. A master mix may then be created composing of sterile water,
BigDye® Terminators and 5x Seq Rxn Buffer, of which 16uL is to be dispensed into
each reaction well. It is strongly recommended that use of this alternative procedure is
validated by the user prior to implementation.
4.5. Add 2µL of purified PCR product to each appropriate well.
NOTE: Care must be taken to prevent cross-contamination of sequence reactions.
4.6. Seal the reaction wells, mix gently and centrifuge briefly to ensure that the contents
are located at the base of each reaction well.
4.7. Place the reaction wells into a thermal cycler and run according to the following
profile:
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Number of cycles
Temperature and time
25
96°C – 10sec
50°C – 5sec
60°C – 2min
1
4°C - hold
4.8. Once the program is complete, remove the reaction wells/plate from the thermal
cycler and either proceed directly to purification of the reaction products or store in
the dark at 4C until required. It is recommended that samples are purified and run
on the DNA sequencer within 24 hours.
5. Purification of Sequencing Reaction Products
NOTE: Purification of the reaction products may be carried out by procedures other than the
ethanol precipitation method described here. It is strongly recommended that users validate
these procedures before proceeding.
5.1. Briefly centrifuge the reaction wells/plates before proceeding. If reusable lids/caps
have been used during thermal cycling, label the lids/caps to avoid crosscontamination.
5.2. Carefully remove the seals.
5.3. To each reaction well add 5µL of 125mM EDTA, pH8.0. Ensure that the EDTA
reaches the base of the reaction well.
5.4. Add 60 µL of 100% ethanol to each reaction well. Seal the wells/plate and vortex
briefly but thoroughly to ensure thorough mixing.
5.5. Pellet the extension products by centrifuging at 2000g for 45 minutes.
IMMEDIATELY PROCEED TO THE NEXT STEP. If this is not possible, recentrifuge for an additional 10 minutes before proceeding.
5.6. Remove the seals to the reaction wells and discard the supernatant by inverting the
reaction wells onto paper towel or tissues.
5.7. Place the inverted reaction wells and paper towel or tissue into the centrifuge.
Centrifuge at 350g for 1 minute to remove any residual supernatant.
5.8. Remove the reaction wells from the centrifuge and place in an upright position on the
work bench. Discard the paper towel or tissues.
5.9. Prepare fresh solution of 80% ethanol with absolute ethanol and sterile water.
5.10. Add 60µL of 80% ethanol to each well. Reseal the wells and vortex briefly.
5.11. Spin at 2000g for 5 minutes.
5.12. Repeat steps 5.6 and 5.7.
5.13. Remove the reaction wells from the centrifuge and discard the paper towel. Reseal
the reaction wells and proceed to the denaturation step. Otherwise store at -20C in
the dark. It is recommended that the extension products are run on the DNA
sequencer within 24 hours of setting up the sequencing reactions.
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For Research Use Only
6. Denaturation & Electrophoresis of Sequencing Reaction Products
NOTE: The procedure for the denaturation of extension products in Hi-Di™ Formamide
described here may not be necessary if purification procedures other than the ethanol
precipitation have been used. It is strongly recommended that users validate alternative
procedures before proceeding.
6.1. Add 12µL of Hi-Di™ Formamide to each reaction well. Vortex and centrifuge the
wells/plate briefly.
6.2. Incubate the reaction wells at 98C for 5 minutes. Following incubation, ensure that
the reaction wells are cooled quickly to room temperature (e.g. place on ice or use
the thermal cycler to perform the denaturation and cooling steps) before being placed
on the sequencer. If it is not possible to run the plates immediately, store at 4C until
required.
NOTE: Ensure that there are no air bubbles in the reaction wells. These can enter and damage
the capillary.
6.3. Load the reaction wells/plate onto the automated sequencer and prepare the data
collection file according to the sequencer manufacturer specifications.
6.4. The following instrument parameters have been validated by the manufacturer using
Big Dye® Terminator Sequencing Kit v3.1 and POP-7™. These parameters may
require user validation for other polymers, sequencing chemistries and instruments.
Please refer to the appropriate instrument user’s manual for detailed instructions and
guidance (e.g. ensure that the dye set setting is appropriate for the chemistry used,
for example v1.1 Big Dye® Terminator sequencing chemistry will require a different
dye set).
Parameter
Setting
Dye set
Z_BigDyeV3
Mobility file
KB_3730_POP7_BDTV3
Basecaller
KB.bcp
Run Module
Regular FastSeq50_POP7
Injection time
15 sec
Run time
3000 sec
6.5. Use the instrument’s data collection software to process the raw collected data and
create the sequence files. Please refer to the appropriate instrument user’s manual for
detailed instructions and guidance.
7. Editing and analysis of electropherograms
The SBT Resolver™ kits were developed and validated using the Assign™ SBT and
Assign™ ATF software developed by Conexio Genomics Pty Ltd. For more details please
refer to the Conexio Genomics website (http://www.conexio-genomics.com).
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For Research Use Only
Performance Characteristics
Conexio Genomics Pty Ltd’s SBT Resolver™ kits are locus specific assays. Sequence
analysis of PCR and sequencing primer sites and performance evaluation has not identified
alleles that are not amplified through the recommended use of these kits.
In most instances the use of the sequencing primers incorporated in each kit will produce a
HLA typing for most samples without the need for further resolution. In those instances
where heterozygous ambiguities remain, the use of resolving sequencing primers (such as
SBT Resolver™ HARPS®) is recommended.
It should be noted that mutations at amplification or sequencing primer sites are possible and
may result in allele drop-out. Samples that suggest a homozygous typing result must be
confirmed by alternative procedures.
Limitations and Cautions

It is strongly recommended that these kits are validated by the user prior to
implementation in the laboratory using samples whose HLA type has been determined by
other molecular based procedures. In particular, any deviations from this procedure (e.g.
the use of alternative PCR or DNA sequencing purification procedures) must be validated
by the user prior to implementation.

These kits have been validated using panels of samples whose genotypes cover a broad
range of alleles. However it should be noted that rare alleles and alleles with
polymorphisms in amplification and sequencing primer sites may be encountered and
these may not be amplified or sequenced.

The nature of HLA sequence based typing is such that factors other than the PCR mix
may result in preferential amplification or allele drop out. As a consequence, apparent
homozygous typing results should be confirmed using alternative methods and/or family
genotyping.

A positive control (human DNA) and negative control (sterile water) must be included on
every PCR run. The positive control must produce a PCR product of the appropriate size
depending on the locus amplified and the resultant sequence must be in concordance with
the sample’s genotype. There must be no PCR products in the negative template control
for each experiment. If a band is evident contamination may have occurred at some level
and the run must be repeated.

Occasionally there may be additional, fainter PCR products evident. These additional
bands do not interfere with sequence results or quality.
License
The SBT Resolver™ kits contain GoTaq® Hot Start Polymerase (DNA POL) which is
manufactured by Promega Corporation for distribution by Conexio Genomics Pty Ltd.
Licensed to Promega under U.S. Patent Nos. 5,338,671 and 5,587,287 and their
corresponding foreign patents.
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For Research Use Only
Troubleshooting
Problem
Possible cause(s)
Solution
No or weak PCR product
Poor quality DNA
Assess DNA quality by gel
electrophoresis. Intact DNA
should be approx 3kb with
little or no evidence of
smearing on gel. Re-extract
DNA and repeat PCR where
possible.
Check concentration of DNA
is between 20-100ng/L. Reextract DNA and repeat PCR
where possible.
Avoid the use of whole blood
specimens containing heparin.
Re-extract DNA and repeat
PCR where possible.
Repeat
PCR.
Ensure
mastermix components are
added and mixed sufficiently
by vortexing.
Check the thermal cycling run
parameters.
Check the run history to ensure
that the run was not terminated
prematurely.
Ensure that the thermal cycler
is operating according to
manufacturer’s specifications
and is regularly maintained.
Submerge the gel in a staining
bath containing 1X TBE with
0.5mg/mL ethidium bromide.
Destain in 1X TBE before
taking gel image.
Ensure ethidium bromide is
added to gel prior to pouring.
Check that the appropriate kit
is used.
Check the thermal cycle
parameters.
Check the negative control for
evidence of contamination.
Decontaminate work area and
repeat PCR.
Repeat PCR to identify source
of contamination. Consider
using a fresh kit.
If the genomic DNA of a
sample
appears
to
be
contaminated, re-extract or
Insufficient quantity of DNA
added to PCR.
Presence of PCR inhibitors in
genomic DNA
DNA polymerase not added to
the mastermix or insufficient
mixing of mastermix prior to
addition to samples.
Thermal cycling problems
No ethidium bromide added to
the gel.
Incorrect band sizes
Incorrect kit used
Incorrect
thermal
program used.
PCR contamination
Page 13 of 16
cycling
For Research Use Only
Weak signal intensity of
electropherograms
Weak PCR product
Insufficient reaction products
applied to sequencer
Problems during purification of
sequencer products
Signal intensity is too
high (Presence of high
fluorescent peaks –
artefacts)
Too much PCR product
Too much reaction products
applied to sequencer.
Noisy baseline (high
background)
Contaminated PCR product
Amplification of closely related
HLA genes
Poor PCR purification
Contaminated
reactions
sequencing
Contaminated sequencing
primer
Contaminated dye terminator
mix or sequencing buffer
Poor purification of sequencing
products.
Page 14 of 16
obtain an alternative source of
DNA.
Check gel image. Sequencing
weak PCR bands is NOT
recommended as the sequence
quality may be insufficient for
SBT.
Consider using a lower
dilution factor (eg 1:2, 1:3)
after PCR purification.
Check sequencer parameters.
Injection time and voltage may
need to be increased.
Use extreme care when
discarding the supernatant as it
may dislodge the pellet.
Check the gel image. Consider
using a higher dilution factor
following PCR purification.
Check the amount of DNA
polymerase used in the PCR.
Check instrument parameters.
Consider
reducing
the
injection time and voltage.
Refer to corrective actions
listed above.
Check
thermal
cycling
parameters. Consider adjusting
the parameters if an alternative
thermal cycler is used.
Ensure ExoSAP-IT® treatment
is undertaken according to
kit’s user instructions.
Ensure that the PCR mixture is
mixed
thoroughly
with
ExoSAP-IT®.
Ensure that all steps are taken
to
prevent
cross
contamination. Change pipette
tips wherever possible. Add
liquids at the top of the
reaction
wells.
Prevent
aerosols.
Check sequence quality of the
other sequencing primers and
other samples using the same
primer.
Repeat sequencing with fresh
aliquot of reagents.
Repeat sequencing and ensure
that purification is undertaken
according to manufacturer’s
instructions.
For Research Use Only
Presence of Dye blobs
Poor purification of sequencing Purify products according to
products
kit instructions.
Ensure products are washed
sufficiently with 80% ethanol.
Related Products
Product code: CGX0036+
XH-PD1.1-2(20)
XH-PD1.1-2(50)
SBT Resolver™ HLA-A kit (20 and 50 tests)
BS-PD2.1-2(20)
BS-PD2.1-2(50)
SBT Resolver™ HLA-B kit (20 and 50 tests)
HH-PD3.2-2(20)
HH-PD3.2-2(50)
SBT Resolver™ HLA-C kit (20 and 50 tests)
PQ-PD6.2-2(20)
PQ-PD6.2-2(50)
SBT Resolver™ HLA-DQB1 kit (20 and 50 tests)
HH-PD10.1(20)
HH-PD10.1(50)
SBT Resolver™ HLA-DPB1 kit (20 and 50 tests)
AN-PD11.0-0(20)
AN-PD11.0-0(50)
SBT Resolver™ HLA-DRB3 kit (20 and 50 tests)
AN-PD12.0-0(20)
AN-PD12.0-0(50)
SBT Resolver™ HLA-DRB4 kit (20 and 50 tests)
AN-PD13.0-0(20)
AN-PD13.0-0(50)
SBT Resolver™ HLA-DRB5 kit (20 and 50 tests)
LC-PD2.9(20)
LC-PD2.9(50)
SBT Resolver™ HLA-B57 kit (20 and 50 tests)
Product codes:
C1-TT98-F(20)
C1-AC98-F(20)
C1-TC98-F(20)
C1-TA98-F(20)
C1-CA102-F(20)
C1-CT102-F(20)
C1-CC102-F(20)
C1-AG203-F(20)
C1-GT240-F(20)
C1-TT368-F(20)
C1-GG307-R(20)
C1-GG363-AF(20)
C1-TA363-F(20)
C1-AT362-F(20)
C1-AC497-F(20)
C1-TA368-F(20)
C1-GT355-R(20)
C1-GG362-R(20)
C1-CT423-F(20)
C1-CG570-R(20)
C1-BTA-F(20)
C1-BCG-F(20)
C1-CC144-F(20)
C1-AC206-F(20)
C1-GC209-F(20)
C1-GA206-F(20)
C1-CG319-F(20)
C1-CA309-R(20)
C1-GAT309-R(20)
C1-GAA309-R(20)
C1-AG360-F(20)
C1-GC363-F(20)
C1-GG363-BF(20)
C1-TA420-F(20)
C1-AC362-F(20)
C1-CC486-F(20)
C1-CT559-R(20)
C1-GA559-R(20)
C1-AC559-R(20)
C1-GG572-R(20)
C1-CG572-R(20)
C1-GAG601-R(20)
C1-CT97-F(20)
C1-CT112-F(20)
C1-CG134-F(20)
C1-AG270-F(20)
C1-AC302-R(20)
C1-GC302-R(20)
C1-CG343-F(20)
C1-CA343-F(20)
C1-GA361-F(20)
C1-TG539-R(20)
C1-GG539-R(20)
C1-AA601-R(20)
RB-01-F(20)
RB-04-F(20)
RB-09-F(20)
RB-15-F(20)
RB-52-F(20)
Page 15 of 16
For Research Use Only
RB-GG125-F(20)
RB-AA197-F(20)
RB-TT197-F(20)
RB-GT196-F(20)
RB-GA196-F(20)
RB-TA164-F(20)
RB-TT227-F(20)
RB-AT258-F(20)
RB-GC258-F(20)
RB-CT257-R(20)
RB-AT257-R(20)
RB-TT321-R(20)
RB-GT344-R(20)
RB-TG344-R(20)
QB-TA173-F(20)
QB-CT173-F(20)
QB-TA185-F(20)
QB-CG353-R(20)
QB-GG353-R(20)
PB-AT251-R(20)
PB-GT313-R(20)
PB-TAC121-F(20)
PB-GG341-R(20)
PB-GC194-F(20)
PB-AG341-R(20)
General Purpose Laboratory Reagents
MgCl2 – 1.0(50)
MgCl2 - 1.0(3000)
2mM MgCl2
SEQ BUF – 2.0(400)
SEQ BUF – 2.0(5000)
5x Seq Rxn Buffer
EDTA – 3.0(200)
EDTA – 3.0(5000)
125mM EDTA, pH8.0
Please contact your local distributor for further details.
Support and Contact Details
Conexio Genomics Pty Ltd
8/31 Pakenham St
Fremantle 6160
Western Australia
Tel: +61-422-863-227
email: support@conexio-genomics.com
Skype: conexiocgx
Website: www.conexio-genomics.com
Or your local distributor
For ordering details, please refer to the Olerup website (http://www.olerup.com).
Conexio and HARPS are trademarks of Conexio 4 Pty Ltd. HARPS® is a registered trademark
in some territories.
Page 16 of 16
For Research Use Only
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