HLA-SuBiTo - the sequencing system
Simple and consistent setup for all HLA-SuBiTo kits
- To separate as many heterozygous allele combinations in the first step, 2 parallel amplifications are performed
- gel electrophoresis
- Purification of PCR products with ExoSAP-IT TM
- 6 sequencing reactions per HLA class I locus sequencing
- 2-3 sequencing reactions per HLA class II locus sequencing
- The combination of forward and reverse reactions occurs depending on the positive PCR amplification
- Loading the capillary sequencer
- Analysis with the HLA-HiType software
Saves 3 hours of assay time with kit enhancements
- New G1 / G2 mix chemistry allows shorter first PCR run time of only 1 h 45 min
- Shorter cycle sequencing PCR protocol lasts only 1 h 10 min
- Alternative purification protocol saves 30 minutes
Kit components
- All components for the amplification incl. IT-Taq polymerase
- Exo-SAP-IT TM for the purification of PCR products
- Genort- and Expon-specific sequencing mixes incl. BigDye ® chemistry
- IT Prec buffer for precipitation of sequencing products
- Two second level sequencing mixes for the resolution of frequent ambiguities.
item number | Surname | Test / plate |
---|---|---|
RUO | Emergency for use in diagnostic procedures - USA and Canada | |
002 090 025 | HLA-SuBiTo A | 25 |
002 090 025/1 | HLA-SuBiTo A without BDT's | 25 |
002 091 025 | HLA-SuBiTo B | 25 |
002 091 025/1 | HLA-SuBiTo B without BDT's | 25 |
002 092 025 | HLA-SuBiTo C | 25 |
002 092 025/1 | HLA-SuBiTo C without BDT's | 25 |
002 093 025 | HLA-SuBiTo DRB1 | 25 |
002 093 025/1 | HLA-SuBiTo DRB1 without BDT's | 25 |
002 094 025 | HLA-SuBiTo DQB1 | 25 |
002 094 025/1 | HLA-SuBiTo DQB1 without BDT's | 25 |
002 095 025 | HLA-SuBiTo DPB1 | 25 |
002 095 025/1 | HLA-SuBiTo DPB1 without BDT's | 25 |
002 096 025 | HLA-SuBiTo ABCDRDQ | 25 |
002 096 025/1 | HLA-SuBiTo ABCDRDQ without BDT's | 25 |
Second level sequencing
Due to the HLA-SuBiTo strategy, which separates HLA allele groups with two initial PCR amplifications, the common cis / trans ambiguities are generally greatly reduced compared to single-amplification strategies. In the event that HLA allele groups are not separated, possible ambiguities can be resolved with our ready-to-use second-level sequencing mixes and the existing standard kit PCR product.
A total of 16 different second-level sequencing mixes are currently available for HLA class I and six each for DRB1 * and DQB1.
The HLA-HiType software shows in ambiguities, with which second level sequencing mix these can be resolved.