Table 1.
Primers Used in PCR Amplification and Sequencing of CFTR Exon 9 and its Intronic Boundaries
| PCR Product | Primers names | Primer sequence (forward primer first) | Amplicon size (bp) | Annealing temp. (°C) |
|---|---|---|---|---|
| A | CF9.6 | 5′-TGGGGAATTATTTGAGAAAGC-3′ | 285 | 50 |
| CF9 + 58 | 5′-CCTTCCAGCACTACAAACTAGAAA-3′ | |||
| B | CF9-130 | 5′-ACAGTGTAATGGATCATGGGC-3′ | 395 | 55 |
| CF9 + 58 | 5′-CCTTCCAGCACTACAAACTAGAAA-3′ | |||
| C*† | CF9-268 | 5′-TGTATACATGTAGTAATTCAGT-3′ | 572 | 55 |
| CF9 + 121 | 5′-ACATTCTCCTAATGCTCATG-3′ | |||
| D*† | CF9-70 | 5′-GTACATAAAACAAGCATCTAT-3′ | 394 | 56 |
| CF9 + 121 | 5′-TGTATACATGTAGTAATTCAGT-3′ | |||
| E*‡ | CF9-70 | 5′-GTACATAAAACAAGCATCTAT-3′ | 710 | 54 |
| CF9 + 437 | 5′-TCTAAATCTATTGAAAATTG-3′ | |||
| F | CF9-184 | 5′-CCTCTAGAAACCGTATGC-3′ | 364 | 54 |
| CF9.181 | 5′-TGCCTGCTCCAGTGGAT-3′ |
*
Indicates specific PCR amplification for CFTR exon 9 and its flanking introns.
†
Two sets of primers were used in which the forward primers are outside the region of similarity, but the reverse primers are inside this region.
‡
Another set of primers was used in which both the forward and reverse primers are outside the region of similarity. This primer design allows the selective amplification of the target region, avoiding amplification of any similar sequences.