Table 2. PCR primers and parameters.

Gene	Forward (5′–3′)	Reverse (5′–3′)	Product size (bp)	Number of cycle	Annealing condition
EGFR
Exon 18	TACACCCAGTGGAGAAGCTCC	CCCCACCAGACCATGAGAG	169	30	58°C, 30 s
Exon 19	CAATTGCCAGTTAACGTCTTCC	GGAGATGAGCAGGGTCTAGAG	239	30	58°C, 30 s
Exon 20	CACACTGACGTGCCTCTC	CTTATCTCCCCTCCCCGTA	252	30	56°C, 30 s
Exon 21	AGGGCATGAACTACTTG	CCTCCTTACTTTGCCTCCTTC	167	35	55°C, 30 s
p53
Exon 5	TTCAACTCTGTCTCCTTCCT	CAGCCCTGTCGTCTCTCCAG	248	30	55°C, 30 s
Exon 6	GCCTCTGATTCCTCACTGAT	TTAACCCCTCCTCCCAGAGA	181	30	55°C, 30 s
Exon 7	CTTGCCACAGGTCTCCCCAA	TGTGCAGGGTGGCAAGTGGC	196	30	59°C, 30 s
Exon 8	TTCCTTACTGCCTCTTGCTT	CGCTTCTTGTCCTGCTTGCT	201	30	55°C, 30 s
K-ras	ACTGAATATAAACTTGTGGTAGTTGGACCT	CTGTATCAAAGAATGGTCCTGCACCAGTA	162	30	58°C, 30 s
p16
Unmethylated	TTATTAGAGGGTGGGGTGGATTGT	CAACCCCAAACCACAACCATAA	151	35	60°C, 30 s
Methylated	TTATTAGAGGGTGGGGCGGATCGC	GACCCCGAACCGCGACCGTAA	150	35	65°C, 15 s
RASSF1A
Methylated	GGGTTTTGCGAGAGCGCG	GCTAACAAACGCGAACCG	169	35	64°C, 50 s
APC1A
Methylated	TATTGCGGAGTGCGGGTC	TCGACGAACTCCCGACGA	98	35	62°C, 10 s