苦瓜ISSR-CR反应体系的建立

Optimization of ISSR-PCR Reaction System for Momordica charantia L.

以苦瓜（Momordica charantia L．）为试材，研究了PCR反应体系的主要成分及退火温度、循环次数对苦瓜ISSR扩增结果的影响。结果表明：在20μl的反应体系中，Mg^2＋的用量为1．2—1．6mmol／L，dNTPs浓度为0．2mmol／L，引物的浓度为0．25μmol／L，Taq DNA聚合酶的用量为1u，模板DNA的用量为30～50ng，在46．4—51.7℃的退火温度下35个循环，能得到清晰、多态性高的ISSR带谱。

The factors that affected the ISSR（ inter-simple sequence repeat） result ofMomordica charantia L. were studied. By adjusting Mg^2 ＋ , dNTPs, primer, Taq DNA polymerase and DNA template concentration, annealing temperature and circles were optimized. The results showed that the optimized content consists of Mg^2＋ 1.2 - 1.6 mmol/L, dNTPs 0.2 mmol/L, primer 0.25 μmol/L, Taq DNA polymerase 1 U, DNA template 30 - 50 ng, annealing temperature was 46.4 - 51.7 ℃, circles was 35 in zoul reaction system.