濒危树种格氏栲ISSR-PCR反应体系的建立

以濒危珍稀树种格氏栲为材料,对ISSR反应体系中的各个主要影响因子进行优化筛选.首次建立可用于格氏栲ISSR-PCR分析的反应体系:20μL PCR反应体系中包括1ng·μL-1模板DNA,0.10 mmo1·L-1dNTP,2.0μL 10×PCR buffer (缓冲液),0.2μmol·L-1引物,0.037 5...

Establishment of ISSR-PCR reaction system of endangered plant Castanopsis kawakamii

The major factors in the reaction system of inter simple sequence repeat(ISSR) amplification of Castanopsis kawakamii Hayata,a rare and endangered plant species,was optimized and selected.The optimal ISSR-PCR system in C.kawakamii was established for the first time,that is,20 μL amplification reactions system containing 1 ng · μL-1 template DNA,0.10 mmol · L-1dNTP,2.0 μL 10×PCR buffer,0.2 μmol · μL-1 primer,0.037 5 U · μL-1 Taq polymerase,11.3 μL ddH2O,2.0 mmol · L-1 MgCl2.The amplified procedure was as follows: after a predenaturing of 5 min at 94 ℃,34 cycles were performed with denaturing of 30 s at 94 ℃,annealing of 45 s,extension of 1.5 min at 72 ℃,and a final extension step of 10 min at 72 ℃ and hold at 4 ℃.