骆驼刺ISSR-PCR反应体系的建立

Study on Alhagi sparsifolia Shap.ISSR-PCR Reaction System

用CTAB法提取叶片DNA,然后通过单因素实验,研究了Taq DNA聚合酶浓度、dNTPs浓度、引物浓度和Mg2+浓度4种因素对ISSR-PCR扩增的影响,建立了适合于骆驼刺的ISSR-PCR反应体系,其为25μL总体积中含1×PCR反应缓冲液(无Mg2+)、1.5U Taq DNA聚合酶、0.6 mmol/L dNTPs、0.6μmol/L引物、1.5 mmol/L MgCl2。

Using the DNA extracted from Alhagi sparsifolia leaf by CTAB method,effect of four factors of the ISSR-PCR reaction system incluting the amount of Taq DNA polymerase,concentration of dNTPs,primers and Mg2＋ was studied by single-factor experiment.The suitable ISSR-PCR reaction system contained l×PCR reaction buffer,1.5 U Taq DNA polymerase,0.6 mmol/L dNTPs,0.6 μmol/L primer,1.5 mmol/L MgCl2 in the 25 μL total volume.