亚麻ISSR的反应体系优化及引物筛选

使用引物U853对亚麻ISSR反应条件进行优化筛选。结果表明:20μL反应体系中,10×Buffer 2μL,Mg2+2.5 mmol/L,Taq酶1.5 U,基因组DNA 50 ng,dNTP 0.25 mmol/L,引物0.5μmol/L。PCR反应程序为:94℃4 min;94℃40 s,55℃1 min,72℃90 s,40个循环;72℃10 min。在该条件下,使用60条ISSR引物对3个有代表性的亚麻(原2003-84、K-6542、SXY8)DNA样品扩增,筛选出13条多态性好,重复性高的引物。

Optimizing the ISSR Reaction System of Flax and Screening Primers

The ISSR reaction conditons of flax were optimized as follows:in 20μL reaction system,it contained 10×Buffer 2μL,Mg2+ 2.5mmol/L,Taq enzyme 1.5U,Genome DNA 50ng,dNTP 0.25mmol/L and ISSR primer 0.5μmol/L.The process of PCR were 94℃ 4min,40 cycles(94℃ 40s,55℃ 1min,72℃ 90s),72℃ 10min.Under this condition 3 typical flax varieties(Yuan 2003-84、K-6542、SXY8) were applified by 60 primers,from which 13 primers were polymorphism and repeatability.This research made the foundation for the molecular identification of flax.