芒果DNA提取方法比较及ISSR反应体系的优化

Comparison of Genomic DNA Extraction Methods and Optimization of ISSR-PCR Reaction Systems for Mango(Mangifera indica L.)

为从芒果幼叶中提取高质量的核总DNA,比较了5种DNA提取方法提取芒果叶片核DNA的效果,结果表明:改良CTAB法1提取的DNA A260/A280值最好,ISSR-PCR扩增效果最佳,是有效提取芒果基因组DNA的方法。为得到最佳的芒果ISSR-PCR反应体系,以(ATG)6为引物,采用单因素实验法,优化了ISSR-PCR反应体系:在总体积25μl的反应体系中,含1×反应缓冲液,0.20mmol.L-1dNTPs,0.20μmol.L-1引物,0.60 UTaqDNA聚合酶,30-50 ng DNA模板,不足体积用无菌超纯水补足。

To get high- quality DNA from young leaves of mango （Mangifera indica L. ）, five different DNA- extracting methods were compared in this paper.The results showed that：The improved CTABI was the best method since the DNA extracted by it had the best A260/A280 ratio and the best ISSR- PCR results. Then the ISSR- PCR reaction system was optimized with primer （ATG）6 by the single factor experiment.The system was：1 × Taq DNA polymerase reaction buffer, 0.6U Taq DNA polymerase,0.2mmol·L^-1 dNTPs, 0.2μmol·L^- 1 primer and 30- 50ng genomic DNA with total 25μl reaction solution.