Table 2 Primer sequences, mixes, and their specificities in PCR-SSP assay
| Primers | Sequences | Annealing temperature (°C) | Primer mixes and concentrations | Amplicon size (bp) |
|---|---|---|---|---|
|
HPA-1a HPA-1b common |
5ʹ-ACTTACAGGCCCTGCCTCT-3ʹ 5ʹ-ACTTACAGGCCCTGCCTCC-3ʹ 5ʹ-AGCCGGAGTGCAATCCTCTG-3ʹ |
62 64 66 |
HPA-1a + common, 0.5 µM HPA-1b + common, 0.5 µM | 196 |
|
HPA-2a HPA-2b common |
5ʹ-CCCCCAGGGCTCCTGAC-3ʹ 5ʹ-GCCCCCAGGGCTCCTGAT-3ʹ 5ʹ-GCCAGCGACGAAAATAGAGG-3ʹ |
64 62 62 |
HPA-2a + common, 0.5 µM HPA-2b + common, 0.5 µM | 241 |
|
HPA-3a HPA-3b common |
5ʹ-GGGGGAGGGGCTGGGGA-3ʹ 5ʹ-GGGGGAGGGGCTGGGGC-3ʹ 5ʹ-GACCTGCTCTACATCCTGGA-3ʹ |
64 66 60 |
HPA-3a + common, 0.5 µM HPA-3b + common, 0.5 µM | 230 |
|
HPA-4a HPA-4b common |
5ʹ-GCTGGCCACCCAGATGCG-3ʹ 5ʹ-AGCTGGCCACCCAGATGCA-3ʹ 5ʹ-GCTGTCCTGGCGTCTGGAG-3ʹ |
62 60 62 |
HPA-4a + common, 0.5 µM HPA-4b + common, 0.5 µM | 158 |
|
HPA-5a HPA-5b common |
5ʹ-AGTCTACCTGTTTACTATCAAAG -3ʹ 5ʹ-AGTCTACCTGTTTACTATCAAAA -3ʹ 5ʹ-CTCTCATGGAAAATGGCAGTA-3ʹ |
62 60 62 |
HPA-5a + common, 2 µM HPA-5b + common, 2 µM | 249 |
|
HPA-6a HPA-6b common |
5ʹ-GACGAGTGCAGCCCCCG-3ʹ 5ʹ-GGACGAGTGCAGCCCCCA-3ʹ 5ʹ-TAGCGGACACAGGAGAAGTC-3ʹ |
60 62 62 |
HPA-6a + common, 0.5 µM HPA-6b + common, 0.5 µM | 163 |
|
HPA-9a HPA-9b common |
5ʹ-GGGCAGCCCCCAGTCCAC-3ʹ 5ʹ-GGGCAGCCCCCAGTCCAT-3ʹ 5ʹ-GACCTGCTCTACATCCTGGA-3ʹ |
64 62 62 |
HPA-9a + common, 0.5 µM HPA-9b + common, 0.5 µM | 212 |
|
HPA-15a HPA-15b common |
5ʹ-TTCAAATTCTTGGTAAATCCTGT -3ʹ 5ʹ-TTCAAATTCTTGGTAAATCCTGG -3ʹ 5ʹ-ATGACCTTATGATGACCTATTC-3ʹ |
60 62 60 |
HPA-15a + common, 2 µM HPA-15b + common, 2 µM | 225 |
|
HGH-F HGH-R |
5ʹ-GCCTTCCCAACCATTCCCTTA-3ʹ 5ʹ-TCACGGATTTCTGTTGTGTTTC-3ʹ |
64 62 | HGH (F + R), 0.2 µM | 429 |
|
DRα-F DRα-R |
5ʹ-GAGGTAACTGTGCTCACGAACAGC-3ʹ 5ʹ-CACGTTCTCTGTAGTCTCTGGG-3ʹ |
74 68 |
DRα (F + R), 0.2 µM for HPA-5 DRα (F + R), 0.1 µM for HPA-15 | 607 |