Table 1.

Primers and PCR set up for pyrosequencing

Gene	Exon	Codon	Primer	Tm for PCR
Ras	Exon 2	12 and 13	F: 5′-GGCCTGCTGAAAATGACTG-3′ R: 5′-(BioTEG) GCTGTATCGTCAAGGCACTCT-3′ S: 5′-CTTGTGGTAGTTGGAGC-3′	58
Ras	Exon 3	61	F: 5′-AATTGATGGAGAAACCTGTCTCTT-3′ R: 5′-(BioTEG) TACTGGTCCCTCATTGCACTGTA-3′ S: 5′-ATATTCTCGACACAGCAG-3′.	65
Braf	Exon 15	600	F: 5′-TGAAGACCTCACAGTAAAAATAGG-3′ R: 5′-(BioTEG) TCCAGACAACTGTTCAAACTGAT-3′ S: 5′-TGATTTTGGTCTAGCTACA-3′	60

Abbreviations: F forward primer, R reverse primer, S sequencing primer, The PCR reactions were set as follows: 95°C for 5 min; 95°C for 15 s, Tm (° C) for 30 s, 72°C for 15 s, repeat steps 2 through 4 (44 times), 72°C for 5 min, and 4°C forever