Primers used for PCR amplification and DNA sequencing
| Primer . | Primer sequence . | Amplified fragment, bp (size) . | Annealing site . | G6PD protein part . |
|---|---|---|---|---|
| PRs | 5′ CTCTGCAGGCCCGCGGAAGCTCGTT 3′ | − 1 336 to − 908 (428) | − 1 336 to − 1 312 from ATG | Promoter region |
| PRas | 5′ CCGCTGCCGCTGCTCTGCATCCCCA 3′ | − 932 to − 908 from ATG | ||
| 1s | 5′ CGGCGATGGGGATGCGGGAGCACTA 3′ | − 987 to − 578 (409) | − 987 to − 963 from ATG | Exon I, no coding sequence |
| 1as | 5′ GCGGAGCGCGGGACAGTACGCTCCT 3′ | Intron I 56 to 32 | ||
| 2s | 5′ AGGACCTCTCAAGAAAGGGGCTAAC 3′ | − 87 to 182 (269) | Intron I − 78 to − 54 | Exon II, Met1-Ser40 |
| 2as | 5′ AAAAGCTGAGGCATGGAGCAGGCAC 3′ | Intron II 63 to 39 | ||
| 3s | 5′ AAGGGTGGAGGATGATGTATGTAGG 3′ | 9 902 to 10 272 (370) | Intron II − 73 to − 49 | Exon III-IV, Gly41-Lys88 |
| 4as | 5′ TGGGGGCTGGTAGAGAGGGCAGAAC 3′ | Intron IV 54 to 30 | ||
| 5s | 5′ CTGGGGCAGAACACACACGGACTCA 3′ | 10 691 to 11 052 (361) | Intron IV − 76 to − 52 | Exon V, Ala89-Ile162 |
| 5as | 5′ ATAGAGTGGTGGGAGCACTGCCTGG 3′ | Intron V 68 to 44 | ||
| 6s | 5′ TGGGAGGGCGTCTGAATGATGCAGC 3′ | 11 569 to 11 876 (307) | Intron V − 87 to − 63 | Exon VI, Gly163-Arg215 |
| 6as | 5′ GGCCAGGTGAGGCTCCTGAGTACCA 3′ | Intron VI 61 to 37 | ||
| 7s | 5′ GGGTGACCCCTCACATGTGGCCCCT 3′ | 11 917 to 12 166 (249) | Intron VI − 74 to − 50 | Exon VII, Phe216-Arg257 |
| 7as | 5′ GGCTCTGCCACCCTGTGCCAGCCT 3′ | Intron VII 49 to 26 | ||
| 8s | 5′ GTTTGGGGTCCCCATGCCCTTGAAC 3′ | 12 405 to 12 628 (223) | Intron VII − 77 to − 53 | Exon VIII, Asp258-Lys288 |
| 8as | 5′ CAGATGGGCCTGCGACAGGGCATGC 3′ | Intron VIII 52 to 28 | ||
| 9s | 5′ TGCACATCTGTGGCCACAGTCATCC 3′ | 12 943 to 13 268 (325) | Intron VIII − 80 to − 56 | Exon IX, Val289-Asp350 |
| 9as | 5′ TGCCCGCACACAGGGCATGCCCAGT 3′ | Intron IX 58 to 34 | ||
| 10s | 5′ GCTCCCACTGAGACACTCACGCACT 3′ | 13 273 to 13 631 (358) | Intron IX − 76 to − 52 | Exon X, Gly351-Lys429 |
| 10as | 5′ GGCCCAGGCCGCCCACCCTCCACA 3′ | Intron X 46 to 23 | ||
| 11s | 5′ CTGGGGCCCGGGGGACTCCACATGGT 3′ | 13 624 to 13 811 (187) | Intron X − 67 to − 41 | Exon XI, Asn430-Ser455 |
| 11as | 5′ ACCCCATAGCCCACAGGTATGCAG 3′ | Intron XI 45 to 22 | ||
| 12s | 5′ GGGGTGGCCTTTGCCCTCCCTCC 3′ | 13 806 to 14 037 (231) | Intron XI − 65 to − 43 | Exon XII, Asp456-Ser486 |
| 12as | 5′ GGCATGAGGTAGCTCCACCCTCAC 3′ | Intron XII 73 to 50 | ||
| 13s | 5′ AGGAAAGGGTGGGGGCTGGGGACAGA 3′ | 13 967 to 14 239 (272) | Intron XII − 94 to − 69 | Exon XIII, Arg487-Leu515 |
| 13as | 5′ GTCAATGGTCCCGGAGTCCTCCCGA 3′ | Exon XIII 177 to 153 | ||
| 3′UTRs | 5′ TTTCCAGTATGAGGGCACCTACAAG 3′ | 14 104 to 14 802 (698) | Exon XIII 43 to 66 | Exon XIII, 3′UTR |
| 3′UTRas | 5′ AAGTGGGTCCTCAGGGAAGCA 3′ | 3′UTR − 42 to − 21 | ||
| CDNAs | 5′ ATATTCATCATCATGGGTGC 3′ | 96 to 262 (166) | Exon II 96 to 115 | Exon II-III, Ser40-Ile80 |
| CDNAas | 5′ GAAGGGCTCACTCTGTTTGC 3′ | Exon III 262 to 243 |
| Primer . | Primer sequence . | Amplified fragment, bp (size) . | Annealing site . | G6PD protein part . |
|---|---|---|---|---|
| PRs | 5′ CTCTGCAGGCCCGCGGAAGCTCGTT 3′ | − 1 336 to − 908 (428) | − 1 336 to − 1 312 from ATG | Promoter region |
| PRas | 5′ CCGCTGCCGCTGCTCTGCATCCCCA 3′ | − 932 to − 908 from ATG | ||
| 1s | 5′ CGGCGATGGGGATGCGGGAGCACTA 3′ | − 987 to − 578 (409) | − 987 to − 963 from ATG | Exon I, no coding sequence |
| 1as | 5′ GCGGAGCGCGGGACAGTACGCTCCT 3′ | Intron I 56 to 32 | ||
| 2s | 5′ AGGACCTCTCAAGAAAGGGGCTAAC 3′ | − 87 to 182 (269) | Intron I − 78 to − 54 | Exon II, Met1-Ser40 |
| 2as | 5′ AAAAGCTGAGGCATGGAGCAGGCAC 3′ | Intron II 63 to 39 | ||
| 3s | 5′ AAGGGTGGAGGATGATGTATGTAGG 3′ | 9 902 to 10 272 (370) | Intron II − 73 to − 49 | Exon III-IV, Gly41-Lys88 |
| 4as | 5′ TGGGGGCTGGTAGAGAGGGCAGAAC 3′ | Intron IV 54 to 30 | ||
| 5s | 5′ CTGGGGCAGAACACACACGGACTCA 3′ | 10 691 to 11 052 (361) | Intron IV − 76 to − 52 | Exon V, Ala89-Ile162 |
| 5as | 5′ ATAGAGTGGTGGGAGCACTGCCTGG 3′ | Intron V 68 to 44 | ||
| 6s | 5′ TGGGAGGGCGTCTGAATGATGCAGC 3′ | 11 569 to 11 876 (307) | Intron V − 87 to − 63 | Exon VI, Gly163-Arg215 |
| 6as | 5′ GGCCAGGTGAGGCTCCTGAGTACCA 3′ | Intron VI 61 to 37 | ||
| 7s | 5′ GGGTGACCCCTCACATGTGGCCCCT 3′ | 11 917 to 12 166 (249) | Intron VI − 74 to − 50 | Exon VII, Phe216-Arg257 |
| 7as | 5′ GGCTCTGCCACCCTGTGCCAGCCT 3′ | Intron VII 49 to 26 | ||
| 8s | 5′ GTTTGGGGTCCCCATGCCCTTGAAC 3′ | 12 405 to 12 628 (223) | Intron VII − 77 to − 53 | Exon VIII, Asp258-Lys288 |
| 8as | 5′ CAGATGGGCCTGCGACAGGGCATGC 3′ | Intron VIII 52 to 28 | ||
| 9s | 5′ TGCACATCTGTGGCCACAGTCATCC 3′ | 12 943 to 13 268 (325) | Intron VIII − 80 to − 56 | Exon IX, Val289-Asp350 |
| 9as | 5′ TGCCCGCACACAGGGCATGCCCAGT 3′ | Intron IX 58 to 34 | ||
| 10s | 5′ GCTCCCACTGAGACACTCACGCACT 3′ | 13 273 to 13 631 (358) | Intron IX − 76 to − 52 | Exon X, Gly351-Lys429 |
| 10as | 5′ GGCCCAGGCCGCCCACCCTCCACA 3′ | Intron X 46 to 23 | ||
| 11s | 5′ CTGGGGCCCGGGGGACTCCACATGGT 3′ | 13 624 to 13 811 (187) | Intron X − 67 to − 41 | Exon XI, Asn430-Ser455 |
| 11as | 5′ ACCCCATAGCCCACAGGTATGCAG 3′ | Intron XI 45 to 22 | ||
| 12s | 5′ GGGGTGGCCTTTGCCCTCCCTCC 3′ | 13 806 to 14 037 (231) | Intron XI − 65 to − 43 | Exon XII, Asp456-Ser486 |
| 12as | 5′ GGCATGAGGTAGCTCCACCCTCAC 3′ | Intron XII 73 to 50 | ||
| 13s | 5′ AGGAAAGGGTGGGGGCTGGGGACAGA 3′ | 13 967 to 14 239 (272) | Intron XII − 94 to − 69 | Exon XIII, Arg487-Leu515 |
| 13as | 5′ GTCAATGGTCCCGGAGTCCTCCCGA 3′ | Exon XIII 177 to 153 | ||
| 3′UTRs | 5′ TTTCCAGTATGAGGGCACCTACAAG 3′ | 14 104 to 14 802 (698) | Exon XIII 43 to 66 | Exon XIII, 3′UTR |
| 3′UTRas | 5′ AAGTGGGTCCTCAGGGAAGCA 3′ | 3′UTR − 42 to − 21 | ||
| CDNAs | 5′ ATATTCATCATCATGGGTGC 3′ | 96 to 262 (166) | Exon II 96 to 115 | Exon II-III, Ser40-Ile80 |
| CDNAas | 5′ GAAGGGCTCACTCTGTTTGC 3′ | Exon III 262 to 243 |