Supplementary Table I.
Primers and conditions of allele-specific primer amplification assays.
| Primers | Sequence 5′ to 3′ | PCR specificity | PCR conditions |
|---|---|---|---|
| 69E3CEAS | 5′ CTGATGACCATCCTCAGGG 3′ | RHCE | 95 °C, 5 min / 10 cycles (94 °C, 20s-67 °C, 30s) / 21 cycles (94 °C, 20s-63 °C, 20s - 72 °C, 20s: 30 cycles) / 72 °C, 5 min / 4 °C inf. 20 ng DNA, 10 μM of each primer, 200 μM of each dNTP (Amersham Biosciences, Buckinghamshire, UK), 1 μL Taq DNA Polymerase (Titanium, Clontech Laboratories, Mountain View, CA, USA) in the appropriate buffer, total reaction volume 50 μL |
| 35I2E3DCES | 5′ CCTTCTCACCCCCAGTATTC 3′ | C340 | |
| 68I2E3VMAYS | 5′ CCTTCTCACCCCCAGTATTT 3′ | T340 | |
| 88TI5’NCS | 5′ ATAGTCCCTCTGCTTCCG 3′ | RHCE/RHD | |
| 89TIEI1AS | 5′ CCAATGAACTCTCACCTTG 3′ | RHCE/RHD | |
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| |||
| 109D-EX3F | 5′ TCGGTGCTGATCTCAGTGGA 3′ | RHD | 94 °C, 15min / 30 cycles (94 °C, 30s-62 °C, 30s-72 °C, 30s) / 72 °C, 10 min / 4 °C inf 50 ng DNA, 12 to 24 μM specific primers, 8 μM GH1 primers, 200 μM of each dNTP (Amersham Biosciences, Buckinghamshire, UK), 0.4 μL HotStarTaq (Qiagen, Hilden, Germany) in the appropriate buffer, total reaction volume 25 μL |
| 118-D-EX3R | 5′ ACTGATGACCATCCTCAGGT 3′ | 455A | |
| 110CE-EX3R | 5′ ACTGATGACCATCCTCAGGG 3′ | 455C | |
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| |||
| 49I5E5CEAS | 5′ TCACCATGCTGATCTTCCT 3′ | RHCE | 95 °C, 15 min / 28 cycles (94 °C, 30s-61 °C, 30s-72 °C, 1 min) / 72 °C, 10 min / 4 °C inf 50 ng DNA, 24 μM of specific primers, 6 μM GH1 primers, 200 μM of each dNTP (Amersham Biosciences, Buckinghamshire, UK), 0.3 μL HotStarTaq (Qiagen, Hilden, Germany) in the appropriate buffer, total reaction volume 25 μL |
| 18EX4CES | 5′ ACTACCACATGAACCTGAG 3′ | RHCE | |
| 17EX4DS | 5′GACTACCACATGAACATGAT 3′ | RN | |
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| |||
| GH1sense | 5′ TGCCTTCCCAACCATTCCCTTA 3′ | GH1 | |
| GH1antisense | 5′CCACTCACGGATTTCTGTTGTGTTTC3′ | GH1 | |