HLA-Sequencing

HLA-SuBiTo - The Sequencing

Same simple and uniform set up for all HLA-SuBiTo kits

  • To reduce ambiguities two amplifications are carried out in parallel in order to separate many heterozygous allele combinations in the first step
  • Gel Electrophoresis
  • Purification of the PCR products using ExoSAP-ITTM
  • 6 sequence reactions per HLA Class I Locus sequence
  • 3-5 sequence reactions per HLA Class II Locus sequence
  • The combination of forward and reverse reactions takes place according to positive PCR amplification
  • Precipitation with IT-Prec-Buffer
  • Loading of the capillary sequencer
  • Analysis with HLA-HiType software

New kit improvements save you 3 hours assay time

  • New chemistry on G1/G2 mixes enable shorter PCR profile for first PCR - takes now only 1 h 45 min
  • Shorter protocol for Cycle Sequencing PCR takes now only 1 h 10 min
  • Shorter precipitation protocol saves 30 minutes

Complete kit contains

  • All components for amplification including IT-Taq Polymerase
  • Exo-SAP-ITTM for purification of the PCR products
  • Gene locus and exon-specific sequencing minxes incl. BigDye® chemicals
  • IT-Prec Buffer precipitation of the sequencing products
  • Second Stage sequencing mixes for resolution of common ambiguities are directly included in the kit
Art. No. Name Test / Plate
002 090 025 HLA SuBiTo A high res 25
002 090 025/1 HLA SuBiTo A high res without BDT's 25
002 091 025 HLA SuBiTo B high res 25
002 091 025/1 HLA SuBiTo B high res without BDT's 25
002 092 025 HLA SuBiTo C high res 25
002 092 025/1 HLA SuBiTo C high res without BDT's 25
002 093 025 HLA SuBiTo DRB1 high res 25
002 093 025/1 HLA SuBiTo DRB1 high res without BDT's 25
002 094 025 HLA SuBiTo DQB1 high res 25
002 094 025/1 HLA SuBiTo DQB1 high res without BDT's 25
002 095 025 HLA SuBiTo DPB1 25
002 095 025/1 HLA SuBiTo DPB1 high res without BDT's 25
002 096 025 HLA SuBiTo ABCDRDQ 25
002 096 025/1 HLA SuBiTo ABCDRDQ high res without BDT's 25

Flyer HLA-SuBiTo


Second Level Sequencing

On the strength of the HLA-SuBiTo two amplification strategy, ambiguous heterozygotes due to cis-trans ambiguities are reduced severely. However in case the groups could not be separated, possible ambiguities can be resolved with the ready-to-use Second Level Sequencing Mixes using the existing PCR product of the standard kit.

There are a total of 16 different Second Level Sequencing Mixes available for HLA class I, 6 different Mixes for HLA-DRB1 and HLA-DQB1, and 4 mixes for HLA-DPB1.

The powerful HLA-HiType Software proposes in the case of ambiguities the ideal Second Level Sequencing Mix for resolution.

Art. No. Name Test / Plate
002 290 001 HLA SuBiTo A Seq2 No1 10
002 290 002 HLA SuBiTo A Seq2 No2 10
002 290 003 HLA SuBiTo A Seq2 No3 10
002 290 004 HLA SuBiTo A Seq2 No4 10
002 290 005 HLA SuBiTo A Seq2 No5 10
002 290 006 HLA SuBiTo A Seq2 No6 10
002 290 013 HLA SuBiTo A Seq2 No13 10
002 290 020 HLA SuBiTo A Seq2 No20 10
002 290 025 HLA SuBiTo A Seq2 No25 10
002 290 027 HLA SuBiTo A Seq2 No27 10
Art. No. Name Test / Plate
002 091 002 HLA-SuBiTo B Seq2 No. 2 10
002 091 006 HLA-SuBiTo B Seq2 No. 6 10
002 091 007 HLA-SuBiTo B Seq2 No. 7 10
002 091 011 HLA-SuBiTo B Seq2 No. 11 10
002 091 013 HLA-SuBiTo B Seq2 No. 13 10
002 091 015 HLA-SuBiTo B Seq2 No. 15 10
002 091 018 HLA-SuBiTo B Seq2 No. 18 10
002 091 021 HLA-SuBiTo B Seq2 No. 21 10
002 091 029 HLA-SuBiTo B Seq2 No. 29 10
002 091 030 HLA-SuBiTo B Seq2 No. 30 10
002 091 031 HLA-SuBiTo B Seq2 No. 31 10
Art. No. Name Test / Plate
002 092 005 HLA-SuBiTo C Seq2 No. 5 10
002 092 006 HLA-SuBiTo C Seq2 No. 6 10
002 092 008 HLA-SuBiTo C Seq2 No. 8 10
002 092 009 HLA-SuBiTo C Seq2 No. 9 10
002 092 010 HLA-SuBiTo C Seq2 No. 10 10
002 092 013 HLA-SuBiTo C Seq2 No. 13 10
002 092 016 HLA-SuBiTo C Seq2 No. 16 10
002 092 017 HLA-SuBiTo C Seq2 No. 17 10
002 092 023 HLA-SuBiTo C Seq2 No. 23 10
002 092 225 HLA-SuBiTo C Seq2 No. 25 10
002 092 028 HLA-SuBiTo C Seq2 No. 28 10
002 092 029 HLA-SuBiTo C Seq2 No. 29 10
Art. No. Name Test / Plate
002 295 001 HLA SuBiTo DPB1 Seq2 No1 10
002 295 002 HLA SuBiTo DPB1 Seq2 No2 10
002 295 003 HLA SuBiTo DPB1 Seq2 No3 10
002 295 004 HLA SuBiTo DPB1 Seq2 No4 10
Art. No. Name Test / Plate
002 294 001 HLA SuBiTo DQB1 Seq2 No1 10
002 294 002 HLA SuBiTo DQB1 Seq2 No2 10
002 294 003 HLA SuBiTo DQB1 Seq2 No3 10
002 294 004 HLA SuBiTo DQB1 Seq2 No4 10
002 294 005 HLA SuBiTo DQB1 Seq2 No5 10
002 294 006 HLA SuBiTo DQB1 Seq2 No6 10
002 294 007 HLA SuBiTo DQB1 Seq2 No7 10
002 294 009 HLA SuBiTo DQB1 Seq2 No9 10

Analysis with HLA-HiType Software

  • Integration of new nomenclature and automated translation in P- and G- groups as well as new NMDP-Codes
  • Fully automated data management, no exhausting selection of *.ab1 Files necessary
  • Automated recognition and dynamic display of hemi- and heterozygous sequences
  • Several manual and automated editing possibilities
  • Diverse alignment options, combined control of electropherogram and alignment
  • High safety due to a defined 4 eyes principle (technical and medical validation)
  • Complete history with a traceability of each step 

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