莲雾SRAP反应体系的优化及其应用

Optimization of SRAP-PCR System and its Application in Wax Apple(Syzygium samarangens)

以莲雾DNA为模板,应用正交设计法对SRAP反应体系中的各个主要影响因素进行了优化筛选。结果表明,20μL反应体系中各组分的最适浓度或用量分别为:1×Buffer,3.0mmol/LMg2+,0.3mmol/LdNTPs,0.4μmol/L引物,0.5UTaqDNA聚合酶和40ng模板DNA。利用该优化体系通过64对SRAP引物组合对7份莲雾进行了SRAP-PCR扩增,证实了该体系具有稳定可靠、重复性好、多态性较强等特点。

The main influential factors of SRAP reaction system were optimized for genomic DNA in wax apple（Syzygium samarangens） by orthogonal design.The results showed that the optimum concentration was 1× Buffer,Mg 2＋ 3.0 mmol/L,dNTPs 0.3 mmol/L,primer 0.4 μmol/L,Taq DNA polymerase 0.5 U,40 ng template DNA with a total volume of 20 μL reaction solution.The molecular identification of 7 varieties of wax apple were achieved by 64 SRAP primer combinations with this optimum system which could provide clear,reliable and repeatable abundant polymorphisms molecular markers.