Table 2. Primer sequences for the amplification of the coding region of DFNB31.
Primer name | Sequence (5′-3′) | Annealing temperature |
---|---|---|
Exon 1 |
F: CAGCAGCCAACTCTTGTGTC |
55 °C |
R: CCAGAAAGGCCAAGTGATTC | ||
Exon 2 |
F: ACTCCCCAAATCCAAGTTCC |
58 °C |
R: CAGAACCAGCCTCTTCTTGC | ||
Exon 3 |
F: CTCCTTGCCAGTCGGATAAG |
55 °C |
R: GAGTGCTGATTGCTCTGCTG | ||
Exon 4 |
F: ATAAGGGGACCCTTGGAATG |
55 °C |
R: TCCCCACTTTTTGGATGAAG | ||
Exon 5 |
F: GTCCGGAGTTTCCTTTACCC |
55 °C |
R: TGGTCTGCTCTGTTCATTGC | ||
Exon 6 |
F: TGGCAATGAACAGAGCAGAC |
58 °C |
R: GGAGGGCTTGTGAAGATGAC | ||
Exon 7 |
F: GACAGGGAAGCAGGAGTGAG |
58 °C |
R: GATTCGAACTCAGGCTGGTC | ||
Exon 8 |
F: CAGCATCTCTGGCAGTTCAG |
58 °C |
R: GGCTGTCATGGAGAGGAGAG | ||
Exon 9 |
F: GTGACAAGCTCTGGCTGATG |
58 °C |
R: TTCAAACTGGGGTCTCCAAC | ||
Exon 10 |
F: GGTCTGGTTGAAAGGACAGG |
58 °C |
R: GGCCTCCAGATTCCTAATCC | ||
Exon 11 |
F: GAGGCTGAGATTGGTCTTGG |
55 °C |
R: CCTAGGTCTGCCCTTGAGTG | ||
Exon 12 | F: CCCTTTCTCAGCATCTCCAG |
58 °C |
R: GTCTGCCTTGTCCTGCTCTC |
Amplification conditions were 95 °C 5 min followed by 35 cycles of 30 s at 95 °C, 30 s at an annealing temperature specific for each exon (column 3) and 30 s at 72 °C.