龙荔基因组DNA的提取及ISSR-PCR体系的建立与优化

Establishment and Optimization of Genomic DNA Extraction and ISSR-PCR Reaction System for Dimocarpus confinis

应用改良的CTAB法提取龙荔基因组DNA,探讨龙荔ISSR-PCR扩增反应体系的建立和优化。结果表明,经过改良的CTAB法提取龙荔基因组DNA效果良好,所提取的DNA纯度较高,符合ISSR-PCR反应的要求;龙荔ISSR-PCR最佳反应体系（反应总体积20μl）：1×Buffer缓冲液（含适量Mg^2＋）,1 UTaqDNA聚合酶,0.20 mmol/L dNTP,0.25μmol/L引物,DNA模板70-80 ng/μl,退火温度53-54℃。

The improved CrAB method was applied to extract genomic DNA of Dimorcarpus confinis to discuss the establishment and optimization of ISSR-PCR reaction system. The results showed that the effects of the extraction genomic DNA by the improved CTAB method was well, the purity of DNA was high,which accorded to the requirement ISSR-PCR Reaction. The optimal reaction system was established for ISSR-PCR of Dimocarpus Confinis ,and 20 μl ISSR-PCR system contained 1 × Tap Buffer,1 U Taq DNA polymerase,0.2 mmol/L dNTP,0. 25 μmol/L primer and 70 -80 ng/μl template DNA.