Table 2

Summary of primer sequences and annealing temperatures for direct sequence

ExonPrimer sequencesTm (°C)Product size (bp)
For DNA templates from frozen samples
18F, 5′-GAGGTGACCCTTGTCTCTGTGT-3′57189
R, 5′-AGCCCAGAGGCCTGTGCCA-3′
19F, 5′-CCAGATCACTGGGCAGCATGTGGCACC-3′65265
R, 5′-AGCAGGGTCTAGAGCAGAGCAGCTGCC-3′
20F, 5′-ACTGACGTGCCTCTCCCTCC-3′57235
R, 5′-CCGTATCTCCCTTCCCTGATT-3′
21F, 5′-ATCTGTCCCTCACAGCAGGGTC-3′57210
R, 5′-GGCTGACCTAAAGCCACCT-3′
22F, 5′-AATTAGGTCCAGAGTGAGTTAAC-3′65251
R, 5′-ACTTGCATGTCAGAGGATATAATG-3′
23F, 5′-CATCAAGAAACAGTAACCAGTAATG-3′65320
R, 5′-AAGGCCTCAGCTGTTTGGCTAAG-3′
24F, 5′-TTGACTGGAAGTGTCGCATCACC-3′65279
R, 5′-CATGTGACAGAACACAGTGACATG-3′
For DNA templates from paraffin-embedded samples
18a*F, 5′-GAGGTGACCCTTGTCTCGTGT-3′56110
R, 5′-ATTCAGTTTCCTTCAAGATCCTC-3′
18b**F, 5′-AGTGGAGAAGCTCCCAACCAAGC-3′65131
R, 5′-AGCCCAGAGGCCTGTGCCA-3′
19F, 5′-AACGTCTTCCTTCTCTCTCTGTCAT-3′56150
R, 5′-CCACACAGCAAAGCAGAAACTC-3′
20a*F, 5′-ACTGACGTGCCTCTCCCTCC-3′56127
R, 5′-AAGGGCATGAGCTGCGTGA-3′
20b**F, 5′-TGCCTCACCTCCACCGT-3′56152
R, 5′-CCGTATCTCCCTTCCCTGATT-3′
21a*F, 5′-ATCTGTCCCTCACAGCAGGGTC-3′56126
R, 5′-TGATCTTGACATGCTGCGGTGTT-3′
21b**F, 5′-AGCCAGGAACGTACTGGTGA-3′56134
R, 5′-GGCTGACCTAAAGCCACCT-3′
  • NOTE. a* and b**, exon 18, 20, and 21 were divided into two parts for PCR amplification and sequenced separately.

    Abbreviations: Tm, annealing temperature; F, forward primer; R, reverse primer.