Fragment | Exon | Primer sequence | Product size (bp) | Annealing (°C) |
---|---|---|---|---|
1 | 2 | F: 5' CTT TGA GAG CCC CGT GAG TG 3' | 718 | 63 |
R: 5' TTG TGA GGT GCG TGG GTG TC 3' | ||||
2 | 3 | F: 5' TCA GAG AGG CTG AAT GTG CTG 3' | 306 | 60 |
R: 5' GCA CAG AGC CCA GAA CTC AC 3' | ||||
3 | 4 & 5 | F: 5' GGT GCG CCT CTT CTG ATA TG 3' | 625 | 60 |
R: 5' GGA AAT GAG TCC TAT GGG CTG 3' | ||||
4 | 6, 7 & 8 | F: 5' TGG GGA GAG AGC CTC AAC TC 3' | 653 | 62 |
R: 5' GCA CAC ACC ACG ATC ATC ATG 3' | ||||
5 | 9 | F: 5' GCT GTA CAC ACG CAT GAT GTC 3' | 269 | 60 |
R: 5' GCC TCT GCT TTC TAA CCC CC 3' | ||||
6 | 10 & 11 | F: 5' TCA GGC TGA GGC TCA GTG GG 3' | 465 | 63 |
R: 5' CAG AGA TGA GGG TGC TAA GTC 3' | ||||
7 | 12 | F: 5' TAA CGC CAG CCC CAC AGA GG 3' | 380 | 60 |
R: 5' TCC GCT TTT CCT CCT CCC TG 3' | ||||
8 | 13 & 14 | F: 5' CAT CAG GTG GCC CAG ACA G 3' | 688 | 62 |
R: 5' GAG AGT CTT GGG TGG GTG G 3' | ||||
9 | 15 | F: 5' ATG CTG GGT GGC TGA GAA G 3' | 390 | 59 |
R: 5' CCA GGC CCA AAT GTT GTC TC 3' | ||||
10 | 16 | F: 5' CCA GCC CCA TCC CAT TCA T 3' | 298 | 59 |
R: 5' GTT CCT CTT TCC GCC ACC T 3' | ||||
11 | 17 | F: 5' AGA TGG AGA GCG TGG TTC C 3' | 287 | 55 |
R: 5' CCC CAG CTC TGC AGT GTG 3' | ||||
12 | 18 & 19 | F: 5' TAG TTA CTG GCA GCC TGG TG 3' | 821 | 57 |
R: 5' ATC CTC TGT TCC TGG CAT CG 3' | ||||
13 | 20 | F: 5' CAC AAT GAC GAC CTC TGA GC 3' | 369 | 56 |
R: 5' GGA ACC AGC CTC CCT CTG 3' | ||||
14* | 17, 18 & 19 | F: 5' AGA TGG AGA GCG TGG TTC C 3' | Wild-type: 1386 Mutant: 848 | 56 |
R: 5' ATC CTC TGT TCC TGG CAT CG 3' |