Table 1. Primers used for amplification of EGFR exons 18-21.

EGFR Exon	Sanger sequencing 5’-3’	NGS 5’-3’ a	Lenght (bp)
Ex18 Fw	CATgTCTggCACTgCTTTCC	ggCTgAggTgACCCTTgTC	184
Ex18 Rv	AgggACCTTACCTTATACACC	gCCTgTgCCAgggACCTTAC
Ex19 Fw	AgCATgTggCACCATCTCAC	AgCATgTggCACCATCTCAC	182
Ex19 Rv	ATgAgAAAAggTgggCCTgA	CCCACACAgCAAAgCAgAAA
Ex20 Fw	AgCCACACTgACgTgCCTCT	AgCCACACTgACgTgCCTCT	208
Ex20 Rv	CCTTATCTCCCCTCCCCgTA	TgCACACACCAgTTgAgCAg
Ex21 Fw	TgCAgAgCTTCTTCCCATgA	CCTCACAgCAgggTCTTCTC	196
Ex21 Rv	gCATgTgTTAAACAATACAgC	CCACCTCCTTACTTTgCCTCCT

^a Primers for 454 next generation sequencing are linked to a sequence of 10 nucleotides (MID – Multiple Identifier), 4 nucleotide of “junction” and 25 nucleotide of “universal sequence”, according to manufacturer’s instruction (not shown in the Table). Ex, Exon; Fw, Forward; Rv, Reverse.