东北刺人参RAPD反应体系的优化

Optimization of RAPD reaction system of Oplpanax elatus Nakai

以东北刺人参试管苗叶片为材料,建立东北刺人参RAPD反应优化体系.结果表明:20.0μL反应体系含250.0μmol/L dNTP,2.0μmol/L MgCl2,1×Buffer,0.3μmol/L引物,10.0 ng DNA模板,1.5 U Taq DNA聚合酶,9.8μL ddH2O;扩增反应程序为:94℃预变性5min,94℃变性45 s,36℃退火1 min,72℃延伸1.5 min,40个循环,最后72℃延伸7 min时获得的RAPD指纹图谱带型清晰、重复性好.

Optimization of RAPDreaction system for Oplpanax elatus Nakai was established with fresh leaves. The results indicated that in 20. 0 μL reaction system, include 250.0μmol/L dNTP, 2.0 mmol/L MgCl2, 1X Buffer, 0.3 μmol./L primer, 10.0 ng templet DNA, 1.5 U Taq DNA polymerase and 9.8/μL ddH2O;amplification procedure was initial denaturation of 5 min at 94 ℃ ;40 cycles of 45 s at 94 ℃, 1 min at 36 ℃ 1.5 min at 72 ℃and finally 7 minute at 72 ℃.