Combined Overlap Extension PCR Method for Improved Site Directed Mutagenesis

Table 2

Details of mutagenesis experiments showing combination of primers in the primer set for the intended base substitutions.


Primer set	Primers	Expected size (bp)	Base pair changes	Joined full length DNA size (bp)	Position of mutation on

1	FGSP1 + RMP1	387	ATa aGg ATt cGc	1925	1002–1005
	RGSP1 + FMP1	1564
2	FGSP1 + RMP2	513	CTa ATa CTg ATt	1925	1125–1128
	RGSP1 + FMP2	1444
3	FGSP1 + RMP3	931	CCc ATa CCg ATt	1925	1545–1548
	RGSP1 + FMP3	1022
4	FGSP1 + RMP4	1019	aGa cGt	1925	1639–1641
	RGSP1 + FMP4	923
5	FGSP1 + RMP5	1166	aGg aGa cGt cGc	1925	1783–1788
	RGSP1 + FMP5	779
6	FGSP1 + RMP6	1441	CTa GAG aGa CGa CTg GAG cGt CGc	1925	2052–2061
	RGSP1 + FMP6	512
7	FGSP1 + RMP7	1649	aGa ATG aGa aGa cGc ATG cGt cGc	1925	2257–2268
	RGSP1 + FMP7	308
8	FGSP2 + RMP8	469	TAt ATc tcc TTc TAc ATt agt TTt	2685	615–624
	RGSP2 + FMP8	2248

Substituted bases are bold and written in lowercase.
of mutation on Stisa2 gene sequence (Accession: AY132997).