Hemi-nested touchdown PCR combined with primer-template mismatch PCR for rapid isolation and sequencing of low molecular weight glutenin subunit gene family from a hexaploid wheat BAC library

Table 2 Position (base in bold) and type of paired base mismatch(es) between primer pair LMWF3/LMWR4 and eight BAC clone templates.

BAC clone (template)	LMWF3 (5'→3') GACAAGTGCCATTGCACAGA	Primer-template mismatch	LMWR4 (5'→3') GCTGTACAACGGCACATTGA	Primer-template mismatch	N-terminal sequence
TaE0503E07	GACAAGTGCCATTGCACAGA	No mismatch	GCTGTACAACGGCACATTGA	No mismatch	METSCISG
TaE1238L16	GACAAGTGCCATTGCACAA A	G:T^a	GCTGTACAACGGCACG TTGA	A:C	METSRVPG
TaE0154F22	GACAAGTGCCATTGCACAA A	G:T	T CTA TACAACGGCACATTGA	G:A, G:T	MENSHIPG
TaE0359D24	A ACAAGTGCCATTGCACAA A	G:T (2×)	T CTGTACAACGGCACATTGA	G:A	IENSHIPG
TaE1157C20	GACAAGTGCCATTGCACAA A	G:T	GCC GTACAC CGGCACG TTGA	T:G, A:G, A:C	METSHIPS
TaE0703A09	GACAAGTGCCATTGCACAA A	G:T	GCC GTACAC CGGCACG TTGA	T:G, A:G, A:C	METSHIPG
TaE0099E23	GACATC TGCCATTGCACAGA	A:A, G:G	GCTGTACAACGGCACATTGA	No mismatch	METSCIPG
TaE0072P19	GACATC TGCCATTGCACAGA	A:A, G:G	GCTGTACAACGGCACATTA A	G:T	METSCIPG

^aThe first base from the primer and the second base from complementary sequence of template.

ISSN: 2730-6844