Table 3. Primers for point mutations in the B-RAF, H, K and N-RAS genes.

Gene	Sequence 5′ ---> 3′	Work concentration (pmol/μl)
BRAF ex15 Forward	CTCATCCTAACACATTTCAAGCC	0.4
BRAF ex15 Reverse	CTATAGTTGAGACCTTCAATGACTTTC	0.4
HRAS ex2 Forward	TGGCTGAGCAGGGCCCTCCT	0.2
HRAS ex2 Reverse	CTGCTGGCACCTGGACGGCGGC	0.2
HRAS ex3 Forward	GGCATGAGAGGTACCAGGGAGA	0.4
HRAS ex3 Reverse	AGGACAGGAGGCCCCTGCCTGGAC	0.4
KRAS ex2 Forward	GGTACTGGTGGAGTATTTGATAGTG	0.2
KRAS ex2 Reverse	CTGACATACTCCCAAGGAAAGTAAAG	0.2
KRAS ex3 Forward	TCCCTTCTCAGGATTCCTACAGG	1.2
KRAS ex3 Reverse	CCCACCTATAATGGTGAATATC	1.2
NRAS ex2 Forward	AGAACCAAATGGAAGGTCAC	0.2
NRAS ex2 Reverse	GTGAGAGACAGGATCAGGTC	0.2
NRAS ex3 Forward	TGAGGGACAAACCAGATAGGC	0.6
NRAS ex3 Reverse	CTGTAGAGGTTAATATCCGCAAATG	0.6

PCR multiplex 2: Oligonucleotides used for the multiplex detection of exon 15 of the BRAF gene, point mutations in codons 12, 13 and 61 (hotspots) of the HRAS gene and in codons 12, 13, 59, 61 of K-NRAS genes. The concentration primers in the mix is reported.