濒危植物刺桫椤RAPD反应体系的优化

Optimal Choice for Reaction System in RAPD Analysis of Alsophila spinulosa

以高盐 SDS法提取濒危植物刺桫椤 ( Alsophila spinulosa)嫩叶总 DNA,进行 RAPD分析 ,分别研究了模板 DNA、引物、 d NTP、 Mg2 + 和 Taq DNA聚合酶用量以及反应体积对反应结果的影响 .筛选并建立了适合于刺桫椤 RAPD扩增的反应体系 :反应体积 1 0 μL,内含 2 ng/ μL模板 DNA,2 .0 mmol/ L Mg Cl2 ,0 .3μmol/ L引物 ,30 0μmol/ L d NTP,0 .5 u Taq DNA聚合酶 .

The DNA templates of Alsophila spinulosa extracted from young leaf were amplified by RAPD.Effects of the content of DNA templates,primers,dNTP,Mg 2+, Taq DNA polymerase and total reaction volume on experimental results were tested and the optimal reaction system of RAPD for A.spinulosa was determined as follows:2ng/μL DNA template,2.0 mmol/L MgCl 2,3.0 μ mol/L primer,300 μ mol/L dNTP,0.5 u Taq DNA polymerase in total 10 μL reaction volume.