Table 3.

List of Primers and Probes Sequences Used in This Study

Target region	PCR round	Primer and probe sequences	Size (bp)
TP53 exon 8	Genomic DNA pre-amplification	F: 5′-GCTTCTCTTTTCCTATCCTG-3′	167
		R: 5′-CTTACCTCGCTTAGTGCT-3′
	Digital PCR	F: 5′-TGGTAATCTACTGGGACG-3′	87
		R: 5′-CGGAGATTCTCTTCCTCT-3′
	Hydrolysis probes	5′-FAM-TGGGAGAGACCGGCGCA-BHQ_1-3′∗
		5′-HEX-TTTGAGGTGCGTGTTTGTGCC-BHQ_1-3′∗
EGFR exon 20	Genomic DNA pre-amplification	F: 5′-GCTGGGCATCTGCCTCACCTCCACCGTGCAACT-3′	91
		R: 5′-GTCTTTGTGTTCCCGGACATAG-3′
	Digital PCR	F: 5′-GCTGGGCATCTGCCTCA-3′	67
		R: 5′-CAGGAGGCAGCCGAAGG-3′
	Hydrolysis probes	5′-FAM-ATGAGTTGCACGGTGGA-BHQ_1-3′∗
		5′-HEX-CTCATCACGCAGCTCATG-BHQ_1-3′∗
		5′-FAM-CTCATCATGCAGCTCATG-BHQ_1-3′†
		5′-HEX-CTCATCACGCAGCTCATG-BHQ_1-3′†

F, forward; R, reverse.

^∗These hydrolysis probes had sequences complementary to the wild-type allele.

^†Hydrolysis probes specific to the mutant (FAM) or wild-type (HEX) allele used for precise quantification of mutational abundances.