Supplementary Table II.
Primers and conditions of exon amplification assays.
| Primers | Sequence 5′ to 3′ | Specificity | PCR assays | PCR conditions |
|---|---|---|---|---|
| 40NC5’DS | 5′ CTCCATAGAGAGGCCAGCACAA 3′ | RHD | RHD exon 1 | 95 °C 5 min: 1 cycle, (95 °C 30s, 63 °C 30s, 72 °C 30s: 30 cycles), 72 °C 5 min: 1 cycle |
| 54I1DAS | 5′ TGCTATTTGCTCCTGTGACCACTT 3′ | RHD | ||
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| 135D2S | 5′TGACGAGTGAAACTCTATCTCGAT 3′ | RHD | RHD exon 2 | 95 °C 10 min: 1 cycle, (92 °C 20s, 64 °C 30s, 68 °C 1 min 30s: 40 cycles), 72 °C 5 min: 1 cycle |
| 136DC2AS | 5′ GGCATGTCTATTTCTCTCTGTCTAAT 3′ | RHD/RHCE | ||
|
| ||||
| 138D3S | 5′ GTCGTCCTGGCTCTCCCTCTCT 3′ | RHD | RHD exon 3 | 95 °C 10 min: 1 cycle, (92 °C 20s, 64 °C 30s, 68 °C 1 min 30s: 40 cycles), 72 °C 5 min: 1 cycle |
| 139DC3AS | 5′ CTTTTCTCCCAGGTCCCTCCT 3′ | RHD/RHCE | ||
|
| ||||
| 148DC4S | 5′GCCGACACTCACTGCTCTTAC 3′ | RHD/RHCE | RHD exon 4 | 95 °C 10 min: 1 cycle (92 °C 20s, 64 °C 30s, 68 °C 1 min 30s: 40 cycles), 72 °C 5 min: 1 cycle |
| 149D4AS | 5′TGAACCTGCTCTGTGAAGTGC 3′ | RHD | ||
|
| ||||
| 17EX4DS | 5′ GACTACCACATGAACATGAT 3′ | RHD | RHD exons 4–5 | 95 °C 5 min: 1 cycle, (95 °C 30s, 60 °C 1 min, 72 °C 1 min 30s: 30 cycles), 72 °C 5 min: 1 cycle |
| 19IN5AS | 5′ AATATGTGTGCTAGTCCTGT 3′ | RHD/RHCE | ||
|
| ||||
| 157DC6S | 5′CAGGGTTGCCTTGTTCCCA 3′ | RHD/RHCE | RHD exon 6 | 95 °C 10 min: 1 cycle, (92 °C 20s, 64 °C 30s, 68 °C 1 min30s: 40 cycles), 72 °C 5 min: 1 cycle |
| 158D6AS | 5′ CTTCAGCCAAAGCAGAGGAGG 3′ | RHD | ||
|
| ||||
| 151D7S | 5′CATCCCCCTTTGGTGGCC 3′ | RHD | RHD exon 7 | 95 °C 10 min: 1 cycle, (92 °C 20s, 60 °C 30s, 68 °C 1 min 30s: 35 cycles), 72 °C 5 min: 1 cycle |
| 152D7AS | 5′ AAGGTAGGGGCTGGACAG 3′ | RHD | ||
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| ||||
| 141D8S | 5′ GGTCAGGAGTTCGAGATCAC 3′ | RHD | RHD exon 8 | 95 °C 5 min: 1 cycle, (95 °C 30s, 64 °C 30s, 68 °C 1 min 30s: 27 cycles), 72 °C 5 min: 1 cycle |
| 142DC8AS | 5′ TGGCAATGGTGGAAGAAAGG 3′ | RHD/RHCE | ||
|
| ||||
| 145D9S | 5′TGCAGTGAGCCGAGGTCAC3′ | RHD | RHD exon 9 | 95 °C 10 min: 1 cycle, (92 °C 20s, 64 °C 30s, 68 °C 1 min 30s: 40 cycles), 72 °C 5 min: 1 cycle |
| 146DC9AS | 5′CACCCGCATGTCAGACTATTTGGC3′ | RHD/RHCE | ||
|
| ||||
| 154DC10S | 5′ CAAGAGATCAAGCCAAAATCAGT 3′ | RHD/RHCE | RHD exon 10 | 95 °C 10 min: 1 cycle, (92 °C 20s, 64 °C 30s, 68 °C 1 min 30s: 40 cycles), 72 °C 5 min: 1 cycle |
| 155D10AS | 5′AGCTTACTGGATGACCACCA 3′ | RHD | ||
|
| ||||
| 41NC5’CES | 5′ CTCCATAGACAGGCCAGCACAG 3′ | RHCE | RHCE exon 1 | 95 °C 5 min: 1 cycle, (95 °C 30s, 63 °C 30s, 72 °C 30s: 30 cycles), 72 °C 5 min: 1 cycle |
| 55I1CEAS | 5′ TGCTATTTGCTCCTGTGACCACTG 3′ | RHCE | ||
|
| ||||
| 73I2DCES | 5′ TCAGTCATCCTGGCTCTCC 3′ | RHD/RHCE | RHD/RHCE exon 3 | 95 °C 5 min: 1 cycle, (95 °C 30s, 60 °C 1 min, 72 °C 1 min 30s: 30 cycles), 72 °C 5 min: 1 cycle |
| 61I3AS | 5′ AGGTCCCTCCTCCAGCAC 3′ | RHD/RHCE | ||
|
| ||||
| 18EX4CES | 5′ ACTACCACATGAACCTGAG 3′ | RHCE | RHCE exons 4–5 | 95 °C 5 min: 1 cycle, (95 °C 30s, 62 °C 30s, 72 °C 30s: 25 cycles), 72 °C 5 min: 1 cycle |
| 19IN5AS | 5′ AATATGTGTGCTAGTCCTGT 3′ | RHD/RHCE | ||
|
| ||||
| 95E5CE3’S | 5′ CCCAAAGGAAGATCAGCAT 3′ | RHCE | RHCE exon 6 | 95 °C 5 min: 1 cycle, (95° C 30s, 68 °C 3 min, 32 cycles), 72 °C 5 min: 1 cycle |
| 96E6I6AS | 5′ TGTCTAGTTTCTTACCGGCA 3′ | RHD/RHCE | ||
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| ||||
| 97IN63’S | 5′ TGTTAGAAATGCTGTTAGACC 3′ | RHD/RHCE | RHCE exon 7 | 95 °C 5 min: 1 cycle, (95 °C 30s, 62 °C 30s, 72 °C 30s: 25 cycles), 72 °C 5 min: 1 cycle |
| 99E7CE3’AS | 5′ CACATGCCATTGCCGTTC 3′ | RHCE | ||