Amplified regions Primer pairs (5’→3’) PCR conditions Length of PCR products
Exon 16 of MLH1 gene f-GGGGTTTTGTTGTGGATTGT r-CTAGTCCTGACCTTGGAACG Initiation denaturation step at 95oC for 10 min, followed by 30 cycles of 95oC for 30 sec, 58oC for 30 sec, 72C for 1 min 30 sec and final elongation step at 72oC for 7 min. 700 bp
Exon 8 of MLH1 gene f-CTCAGCCATGAGACAATAAATC r-GGTTCCCAAATAATGTGATGG 217 bp
Exon 7 of MSH2 gene f-GCTGATTTAGTTGAGACTTACGTGC r-TCTGAATGTGTCCTAAGAGTGAGT 401 bp
Exon 15 of APC gene (codons 967-1386) f-AGTAGTAGTGATGGTTATGGT r-CATCTGCTAAACATGAGTGG 1260 bp
Exon 15 of APC gene (codons 1286–1513) f-GAAATAGGATGTAATCAGACG r-CATTCCCATTGTCATTTTCC 751 bp
Exons 5-6 of the TP53 gene f-TGTTCACTTGTGCCCTGACT r-TTAACCCCTCCTCCCAGAGA 1 cycle of denaturation (2min at 94°C), 20 cycles of denaturation (94°C for 30s), annealing (63°C for 45s, with –0.5°C per cycle), and extension (72°C for 1min), followed by 30 cycles of denaturation (94°C for 30s), annealing (60°C, 45s), extension (72°C for 1min), and a final extension cycle of 72°C for 10min. 467 bp
Exon 7 of the TP53 gene f-CTTGCCACAGGTCTCCCCAA r-AGGGGTCAGAGGCAAGCAGA 237 bp
Exons 8-9 of the TP53 gene f-TTGGGAGTAGATGGAGCCT r-AGTGTTAGACTGGAAACTTT 445 bp
Table 1: The PCR amplification protocols for direct sequencing