TABLE 1.
Primer sequences, binding positions, product sizes, and thermal cycling conditions used in HLA PCR typinga
| Primer | Sequence | Target (relative exon binding position) | PCR parameters
|
Product size (bp) | |
|---|---|---|---|---|---|
| Annealing temp (°C) | No. of cycles | ||||
| HLA#A2F | GTGGATAGAGCAGGAGGGT | HLA-A exon 2 (149-167) | 70 | 7 | 491 |
| HLA#A2R | CCAAGAGCGCAGGTCCTCT | HLA-A exon 3 (110-127) | 65 | 30 | |
| 55 | 3 | ||||
| AL#6 | CGGAATGTGAAGGCCCAG | HLA-A exon 2 (192-209) | 66 | 5 | 447 |
| AL#H | CAAGAGCGCAGGTCCTCT | HLA-A exon 3 (110-127) | 56 | 30 | |
| 50 | 5 | ||||
| β2M#5 | AGATTCAGGTTTACTCACG | β2-M exon 2 (132-150) | 66 | 5 | 264 |
| β2M#6 | TAACTATCTTGGGCTGTGA | β2-M exon 3 (377-395) | 50 | 35 | |
a
All PCRs included 15 min of heat activation of DNA polymerase enzyme at 95°C. Each denaturation and extension phase was 15 s at 94°C and 30 s at 72°C. The annealing temperatures and the cycle numbers are indicated. All exon locations and product sizes are based on sequences with the following accession numbers: HLA-A*0201, AJ575565; HLA-A*68, AJ633570; β2-M, M17987.