SBTexcellerator® reagents

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Instructions, Protocol Sheets, and Certificates

You can find these on our downloads page

Can I buy the primers in the SBTexcellerator kits separately?

Yes, you can order individual primers from the SBTexcellerator® HLA Class I, HLA-DRB1, HLA-DQB1 and HLA-DPB1 Extended kits.
visit our SBTexcellerator® Supplement webpage for more information.

How long are the SBTexcellerator reagents stable at room temperature?

They are stable for over a month and do not require refrigerated transport. For long-term storage however, we recommend storing them at -20°C

How long can a reconstituted primer be stored at -20°C?

Primers are stable for at least 5 months after reconstitution in nuclease-free H2O, when stored at -20°C. Primers can also go through at least 25 freeze/thaw cycles without any negative effects on the results.

When using the SBTexcellerator protocol, when is it OK to stop and for how long can the products be stored?

You can stop the SBTexcellerator protocol:

  1. After amplification PCR
    Store overnight at 15°C in thermocycler or 4°C in fridge. Continue with ExoI/SAP clean-up preferably within 24 hours but definitely within a week after amplification.
  2. After clean-up by ExoI/SAP
    Store overnight at 15°C in thermocycler or 4°C in fridge. Store for 1-2 weeks at 4°C. Store for >2 weeks at -20°C.
  3. After sequencing PCR
    Store for 1-2 days at 15°C in thermocycler or 4°C in fridge.
  4. After clean-up by Sephadex
    Store for 1-2 days at 4°C. Store for >2 days at -20°C. After a longer period of storage it is recommended to denature the reactions by putting them in a thermocycler for 2 min at 98°C and afterwards directly transfer the reactions to a cooling block or ice before running the plate on the sequencing instrument.

IMPORTANT: During storage, always keep your plates sealed to prevent evaporation!

Are the PCR amplification primers for the SBTexcellerator HLA-A kit multiplexed, or is there just a single set of amplification primers yielding one large PCR amplicon?

The PCR primers of the SBTexcellerator HLA-A kit are not multiplexed but are a single set of amplification primers positioned at the UTR regions yielding one large PCR amplicon.

Problems with reading from exon 2 into exon 1 in HLA-B*18 no matter which primer is used?

HLA-B *18 is well known for this. It has an InDel in front of exon 2 making it impossible to read through into exon 1. To read exon 1, you need to use the special exon 1 primers.

How can we solve DQB1 dropouts?

For DQ-02 dropouts

Use more LongRange PCR enzyme (0.8 µl instead of 0.4 µl/rxn).

 

For DQ-03 dropouts

Fresh Q solution usually works. However, if this doesn't work, try 0.8 µl enzyme as above

 

end faq

 
 

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