CFTR primers, PCR and SSCP analysis conditions
| Region amplified | Primer sequence | Anneal temp (°C) | Product size (bp) | Enzyme digest | SSCP conditions % glycerol | |
|---|---|---|---|---|---|---|
| Exon 1 | F 5' - GCC AAA TGA CAT CAC AGC AGG - 3′ | 61 | 211 | None | 3 | |
| R 5′ - TGT CTT TCC GAA GCT CGG TTG - 3′ | ||||||
| Exon 2 | 2i-5 | F 5′ - CCA AAT CTG TAT GGA GAC CA - 3′ | 56 | 203 | None | 3 |
| R 5′ - TCT CTC TTC AAC TAA ACA ATG TAC - 3′ | ||||||
| Exon 3 | 3i-5 | F 5′ - CTT GGG TTA ATC TCC TTG GA - 3′ | 55 | 309 | MboI | 2 |
| 3i-3 | R 5′ - ATT CAC CAG ATT TCG TAG TC - 3′ | |||||
| Exon 4 | 4i-5 | F 5′ - TCA CAT ATG GTA TGA CCC TC - 3′ | 56 | 438 | BslI | 2 |
| 4i-3 | R 5′ - TTG TAC CAG CTC ACT ACC TA - 3′ | |||||
| Exon 5 | F 5′ - GAG AAG ATA GTA AGC TAG ATG AAT - 3′ | 56 | 212 | None | 3 | |
| R 5′ - CAT ATC TCA TCA AAT TTG TTC AGG - 3′ | ||||||
| Exon 6a | F 5′ - GGA AGA TAC AAT GAC ACC TGT TTT - 3′ | 58 | 275 | None | 2 | |
| R 5′ - TAC TAA AGT GGG CTT TTT GAA AAC - 3′ | ||||||
| Exon 6b | F 5′ - CCT TGA GCA GTT CTT AAT AGA TAA - 3′ | 56 | 285 | None | 2 | |
| R 5′ - GGA AGT CTA CCA TGA TAA ACA TAT - 3′ | ||||||
| Exon 7 | F 5′ - CAG AAC TGA AAC TGA CTC GG - 3′ | 58 | 230 | None | 2 | |
| R 5′ - TGC TCC AAG AGA GTC ATA CC - 3′ | ||||||
| Exon 8 | F 5′ - GCA CAA TGA GAG TAT AAA GTA GAT - 3′ | 56 | 223 | None | 3 | |
| R 5′ - AAA CAG TTA GGT GTT TAG AGC AAA - 3′ | ||||||
| Exon 9 (1) | F 5′ - TAA TGG ATC ATG GGC CAT GT - 3′ | 55 | 560 | N/A | N/A | |
| R 5′ - ACA GTG TTG AAT GTG GTG CA - 3′ | ||||||
| Exon 9 (2) | F 5′ - GGG GAA TTA TTT GAG AAA GC - 3′ | 55 | 258 | None | 3 | |
| R 5′ - TTC CAG CAC TAC AAA CTA GAA - 3′ | ||||||
| Exon 10 | 10i-5 | F 5′ - GCA GAG TAC CTG AAA CAG GA - 3′ | 58 | 491 | NsiI/HinfI | 2 |
| 10i-3 | R 5′ - CAT TCA CAG TAG CTT ACC CA - 3′ | |||||
| Exon 11 | 11i-5 | F 5′ - CAA CTG TGG TTA AAG CAA TAG TGT - 3′ | 58 | 425 | HincII | See text |
| 11i-3 | R 5′ - GCA CAG ATT CTG AGT AAC CAT AAT - 3′ | 10 | ||||
| Exon 12 | 12i-5 | F 5′ - GTG AAT CGA TGT GGT GAC CA - 3′ | 60 | 426 | BstNI | 2 |
| 12i-3 | R 5′ - CTG GTT TAG CAT GAG GCG GT - 3′ | |||||
| Exon 13.1 | F 5′ - GAC ATA TTG CAA TAA AGT ATT TAT - 3′ | 53 | 280 | None | 2 | |
| R 5′ - AGT TAG GAT TGA ATT TCT TCT TT - 3′ | ||||||
| Exon 13.2 | F 5′ - TTT AGC TCA AAA CTC ATG GGA - 3′ | 56 | 309 | None | 2 | |
| R 5′ - AGA ATC TGG TAC TAA GGA CAG - 3′ | ||||||
| Exon 13.3 | F 5′ - AAT CCA ATC AAC TCT ATA CGA A - 3′ | 55 | 444 | HincII | 2 | |
| R 5′ - GAT TCT ACT CAA TTG CAT TCT GT - 3′ | ||||||
| Exon 13.4 | F 5′ - CCT GAA CCT GAT GAC ACA C - 3′ | 58 | 239 | None | 2 | |
| R 5′ - GAT TCT ACT CAA TTG CAT TCT GT - 3′ | ||||||
| Exon 14a | F 5′ - GGT GGC ATG AAA CTG TAC TGT CTT - 3′ | 60 | 250 | None | 2 | |
| 14Ai-3 | R 5′ - GTA TAC ATC CCC AAA CTA TCT TAA TTT AAC - 3′ | |||||
| Exon 14b | F 5′ - TGG GAG GAA TAG GTG AAG ATG TTA - 3′ | 60 | 208 | None | 2 | |
| R 5′ - GCT TGG GAG AAA TGA AAC AAA GTG - 3′ | ||||||
| Exon 15 | F 5′ - ATT CAG TAA GTA ACT TTG GCT GCC - 3′ | 60 | 413 | HinfI | 4 | |
| R 5′ - AAG GCA CAT GCC TCT GTG CA - 3′ | ||||||
| Exon 16 | F 5′ - GCG TCT ACT GTG ATC CAA ACT TAG - 3′ | 62 | 289 | RsaI | 3 | |
| R 5′ - TAG ACA GGA CTT CAA CCC TCA ATC - 3′ | ||||||
| Exon 17a | F 5′ - TGA AAA TGT TTA CTC ACC AAC ATG - 3′ | 56 | 254 | None | 2 | |
| R 5′ - AGA ATC TCA AAT AGC TCT TAT AGC - 3′ | ||||||
| Exon 17b | 17Bi-5 | F 5′ - TTC AAA GAA TGG CAC CAG TGT - 3′ | 57 | 463 | HincII | 2 |
| 17Bi-3 | R 5′ - ATA ACC TAT AGA ATG CAG CA - 3′ | |||||
| Exon 18 | F 5′ - ACA TGA GGT TCA TTT ACG TCT TTT - 3′ | 56 | 201 | None | 3 | |
| R 5′ - TAC TCA TAC TTT GTT ACT TGT CTG - 3′ | ||||||
| Exon 19 | 19i-5 | F 5′ - GCC CGA CAA ATA ACC AAG TGA - 3′ | 58 | 454 | HincII | 2 |
| 19i-3 | R 5′ - GCT AAC ACATTG CTT CAG GCT - 3′ | |||||
| Intron 19 | F 5′ AGG CTT CTC AGT GAT CTG TTG - 3′ | 60 | 477 | HphI | See text | |
| R 5′ - GAA TCA TTC AGT GGG TAT AAG CAG - 3′ | 0 | |||||
| Exon 20 | F 5′ - GGT CAG GAT TGA AAG TGT GCA - 3′ | 56 | 473 | HinfI | 2 | |
| R 5′ - CTA TGA GAA AAC TGC ACT GGA - 3′ | ||||||
| Exon 21 | 21i-5 | F 5′ - AAT GTT CAC AAG GGA CTC CA - 3′ | 62 | 477 | DraI | 2 |
| 21i-3 | R 5′ - CAA AAG TAC CTG TTG CTC CA - 3′ | |||||
| Exon 22 | F 5′ - GCT TGA GTG TTT TTA ACT CTG TGG - 3′ | 61 | 266 | None | 2 | |
| R 5′ - GTT CCC ACT GTG CTA TTA AGT AAC - 3′ | ||||||
| Exon 23 | F 5′ - AGC TGA TTG TGC GTA ACG CT - 3′ | 58 | 400 | MboI | 2 | |
| R 5′ - TAA AGC TGG ATG GCT GTA TG - 3′ | ||||||
| Exon 24 | F 5′ - AGA TCT CAC TAA CAG CCA TTT CCC - 3′ | 62 | 263 | None | 3 | |
| R 5′ - CAT GAG CAA ATG TCC CAT GTC AAC - 3′ | ||||||