扁桃基因组DNA提取及RAPD扩增体系的建立

以 13 个扁桃品种幼叶为试材,针对扁桃叶片含有较高的多糖、多酚、色素以及单宁等次生物质的特点,采用改进 SDS 法,在缓冲液中使用 5 mol/L 的 NaCl并在温浴后加入 1/5 体积的 5 mol/L 的 KAc,置于-20℃冰冻 30min,提取到了高质量的基因组 DNA,完全能够用于 RAPD 扩增。同时,分析了模板 DNA 浓度、Taq DNA 聚合酶、dNTPs 浓度对 RAPD 反应体系的影响,建立了一套适合扁桃基因组 RAPD 分析的技术体系。

DNA extraction and the establishment of RAPD amplified for almond cultivars

An improved SDS extraction method was used to obtain high-quality genomic DNA against to rich in polysaccharide, polyphenol, pigment, tannin and other secondary metabolites in blade of 13 cultivars of almond (Amygdalus communis). The method included using 5 mol/L NaCl in reaction solution, adding 1/5 volum Kac, and freezing at -20℃ for 30min. An ideal genomic DNA was gotten and could be used for RAPD. Meanwhile, some factors were optimized, such as concentration of DNA as template, Taq DNA polymerase and dNTPs. A suitable reaction system was also established for RAPD–PCR.