Table 4. PCR conditions for Sanger sequencing and ASLNAqPCR.

Sanger Sequencing
Amplicon	Temperature	Time	Cycles
BRAF (Ex15)	95°C	4′	1
	95°C	30″	40
	53°C	30″	40
	72°C	30″	40
	72°C	10′	1
KRAS (Ex2 and Ex3)a	95°C	4′	1
	95°C	30′	5
	63°C–1°C/cycle	30′	5
	72°C	30″	5
	95°C	30″	35
	58°C	30″	35
	72°C	30″	35
	72°C	10′	1

Ex, exon; ^aFor KRAS amplification a touch-down PCR was performed; ^bPCR for wild type KRAS and all 7 codon 12 and 13 mutations; ^cPCR for wild type BRAF and BRAF V600E; *Plate reading step.