Amplification conditions, primers, and sizes of PCR products for SCN5A coding region. PCR programmes (a-g) are given in Methods. The value following programme (a) indicates the annealing temperature for each PCR amplicon. Primers were taken from Wang et al,22 except those with an asterisk which were redesigned
| Exon | Size (bp) | Primers |
PCR fragment (bp) |
PCR programme |
Special conditions | |||||
| 2 | 324 | F:CCAGAAGCAGGATGAGAA* | 329 | (b) | ||||||
| R:AGAAGGGTAGGCAGGGCT* | ||||||||||
| 3 | 117 | F:AGTCCAAGGGCTCTGAGCCAA | 227 | (b) | ||||||
| R:GGTACTCAGCAGGTATTAACTGCAA | ||||||||||
| 4 | 89 | F:GGTAGCACTGTCCTGGCAGTGAT | 294 | (b) | ||||||
| R:CCTGGACTCAAGTCCCCCTTC | ||||||||||
| 5 | 128 | F:TCACTCCACGTAAGGAACCTG | 324 | (c) | ||||||
| R:ATGTGGACTGCAGGGAGGAAGC | ||||||||||
| 6 | 91 | F:CCTTTCCTCCTCTCACTGTCTGT | 281 | (c) | ||||||
| R:GGTATTCTGGTGACAGGCACATTC | ||||||||||
| 7 | 230 | F:TGACCCCGTGCTTGTTCTTGCCT* | 307 | (a) 64°C | ||||||
| R:GTCTGCGGTCTCACAAAGTCTTC | ||||||||||
| 8 | 63 | F:CCTGGATGCAAGGCGGAAACT* | 180 | (b) | ||||||
| R:GGAGACTCCCCTGGCAGGACAA | ||||||||||
| 9 | 141 | F:GGGAGACAAGTCCAGCCCAGCAA | 262 | (b) | ||||||
| R:AGCCCACACTTGCTGTCCCTTG | ||||||||||
| 10 | 198 | F:ACTTGGAAATGCCCTCACCCAGA | 327 | (c) | 2.0 mmol/l | |||||
| R:CACCTATAGGCACCATCAGTCAG | MgCl2 | |||||||||
| 11 | 178 | F:AAACGTCCGTTCCTCCACTCT | 227 | (b) | ||||||
| R:AACCCACAGCTGGGATTACCATT | ||||||||||
| 12 | 371 | F:GCCAGTGGCTCAAAAGACAGGCT | 280 | (b) | ||||||
| R:CCTGGGCACTGGTCCGGCGCA | ||||||||||
| F:CACCACACATCACTGCTGGTGC | 286 | (b) | ||||||||
| R:GGAACTGCTGATCAGTTTGGGAGA | ||||||||||
| 13 | 132 | F:CCCTTTTCCCCAGCTGACGCAAA | 282 | (b) | ||||||
| R:GTCTAAAGCAGGCCAAGACAAATG | ||||||||||
| 14 | 238 | F:AACTAACTCATTGGCTGTCC* | 295 | (d) | 2.5 mmol/l | |||||
| R:CATTTGAGAGCCAGCTGCAG* | MgCl2 | |||||||||
| 15 | 173 | F:CTTTCCTATCCCAAACAATACCT | 247 | (c) | ||||||
| R:CCCCACCATCCCCCATGCAGT | ||||||||||
| 16 | 350 | F:GAGCCAGAGACCTTCACAAGGTCCCCT | 267 | (a) 64°C | ||||||
| R:TGAAAGAAGTCCATCATGTGCCAGCGA* | ||||||||||
| F:AACTACTCGGAGCTGAGGGA* | 211 | (e) | ||||||||
| R:CCCTTGCCACTTACCACAAG | ||||||||||
| 17 | 440 | F:GGGACTGGATGGCTTGGCATGGT | 306 | (b) | ||||||
| R:CGGGGAGTAGGGGGTGGCAATG | ||||||||||
| F:GCCCAGGGCCAGCTGCCCAGCT | 283 | (c) | ||||||||
| R:CTGTATATGTAGGTGCCTTATACATG | ||||||||||
| 18 | 161 | F: AGGGTCTAAACCCCCAGGGTCA | 280 | (b) | ||||||
| R:CCCAGCTGGCTTCAGGGACAAA | ||||||||||
| 19 | 120 | F:GAGGCCAAAGGCTGCTACTCAG | 245 | (b) | ||||||
| R:CCTGTCCCCTCTGGGTGGAACT | ||||||||||
| 20 | 154 | F:ACAGGCCCTGAGGTGGGCCTGA | 287 | (b) | ||||||
| R:TGACCTGACTTTCCAGCTGGAGA | ||||||||||
| 21 | 173 | F:TCCAGGCTTCATGTCCACCTGTCT | 268 | (b) | ||||||
| R:TCTCCCGCACCGGCAATGGT | ||||||||||
| 22 | 121 | F:AGTGGGGAGCTGTTCCCATCCT | 216 | (b) | ||||||
| R:GGACCGCCTCCCACTCC | ||||||||||
| 23 | 282 | F:AGGAACTGAGCCACTTCTCT* | 343 | (e) | ||||||
| R:AGGAAGGAAGTCCCTTCTCT* | ||||||||||
| 24 | 53 | F:CTCAAGCGAGGTACAGAATTAAATGA | 202 | (b) | ||||||
| R:GGGCTTTCAGATGCAGACACTGAT | ||||||||||
| 25 | 137 | F:GCCTGTCTGATCTCCCTGTGTGA | 265 | (b) | ||||||
| R:AGATGGGACCTGGAGCCTGAGTG* | ||||||||||
| 26 | 104 | F:CCATGCTGGGGCCTCTGAGAAC | 249 | (b) | ||||||
| R:GGCTCTGATGGCTGGCCATGTG | ||||||||||
| 27 | 270 | F:CCCAGCGAGCACTTTCCATTTG | 337 | (b) | ||||||
| R:GCTTCTCCGTCCAGCTGACTTGTA | ||||||||||
| 28 | 3257 | F:ACCAGTAGCCACAGTCTCTGTTG* | 334 | (a) 64°C | ||||||
| R:GAAGAGGCACAGCATGCTGTTGG | ||||||||||
| F:AAGTGGGAGGCTGGCATCGAC | 301 | (b) | ||||||||
| R:GTGCTCTCCTCCGTGGCCACGC | ||||||||||
| F:GAGCCCAGCCGTGGGCATCCT | 309 | (b) | ||||||||
| R:GTCCCCACTCACCATGGGCAG | ||||||||||
| F:CCAACCAGATAAGCCTCATCAACA | 308 | (b) | ||||||||
| R:CCGCCTGCTGACGGAAGAGGA | ||||||||||
| F:TGCTGCAACGCTCTTTGAAGCAT | 346 | (b) | ||||||||
| R:AAAGGCTGCTTTTCAGTGTGTCCT | ||||||||||