A new primer construction technique that effectively increases amplification of rare mutant templates in samples

BMC Biotechnology

Table 2 Primer sequences used in direct PCRs and stuntmer PCRs

Gene	Primer name	Sequence (5′➔ 3′)	Amplicon size
Traditional PCR
EGFR exon 19	Exon 19_Forward	GCAATATCAGCCTTAGGTGCG	323 bps
EGFR exon 19	Exon 19_Reverse	AGCAGCTGCCAGACATGAGA	323 bps
EGFR exon 20	Exon 20_Forward	GAAACTCAAGATCGCATTCATG	365 bps
EGFR exon 20	Exon 20_Reverse	CAAACTCTTGCTATCCCAGGAG	365 bps
EGFR exon 21	Exon 21_Forward	CAGCCATAAGTCCTCGACGTG	399 bps
EGFR exon 21	Exon 21_Reverse	GAGCTCACCCAGAATGTCTGG	399 bps
Stuntmer PCR (see NOTE)
EGFR exon 19 deletion
Forward	*CCCGTCGCT*ATCAAGGAATTAAGAGAAGCAAC-C3-TAAAATTCCCGTCGCT		148 bps
Reverse	AGCAGCTGCCAGACATGAGA		148 bps
EGFR S768
Forward	*TACGTGAT*GGCCAGCGTGGACAACC-C3-CCAGGAAGCCTACGTGAT		277 bps
Reverse	CAAACTCTTGCTATCCCAGGAG
EGFR T790
Forward	*ACCGTGCA*GCTCATCACGCAG-C3-CTCACCTCCACCGTGCA		213 bps
Reverse	CAAACTCTTGCTATCCCAGGAG
EGFR L858
Forward	*GATTTT*GGGCTGGCCAAACTGCTGG-C3-AGCATGTCAAGATCACAGATTTT		203 bps
Reverse	GAGCTCACCCAGAATGTCTGG

The R region: red; E region: blue; linker (C3):grey; overlapping area: highlighted in yellow

ISSN: 1472-6750