Table 1.
Deletion | PCR primers | Fragment size* | Annealing temperature | Sequencing primers |
Promoter – exon1 | F: gtccgaggaaggtcacagaa R: cagcacacaaatgaggtggt |
3.5 kb | 60 | R: cagcacacaaatgaggtggt |
Exon5 | F: gtggtctcggccatccatcc R: tctgcaagccgcctgcaccg |
1.3 kb | 72** | R: tctgcaagccgcctgcaccg |
Exon7 – 8 | F: tcctccttcctctctctggc R: gctgcaggcaggggcgacgc |
3 kb | 63 | R: aaagccaggcacggtggctc |
Exon9 – 14 | F: ggctacaagtgccagtgtga R: agctgacctttagcctgacg |
2 kb | 59 | F: tttttgagacagagtctca R: aaagtccaaaatcaggcc |
Exon 13 – 15 | F: tctccttatccacttgtgtgtctag R: gctttggtcttctctgtctttgaat |
8 kb | 58 | F: tagccaggtgtggtggcagg R: ctgggagtagctaggactgc |
F: forward primer, R: reverse primer
* Fragment size for PCR fragment of the mutant allele
** Two-step PCR, 72°C 1:30 min; 98°C 10sec