| 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 |
