适于AFLP分析的核桃幼叶DNA提取方法

Extraction of DNA for AFLP amplification reaction in teuder walnut leaves

以核桃幼叶为材料,采用改良CTAB法对核桃DNA提取进行了研究。经紫外分光光度计和琼脂糖凝胶电泳检测,在CTAB提取液中添加3%PVP40、50mmol/Lβ-巯基乙醇和10mmol/LNa2S2O5,能得到高纯度的总DNA,OD260/OD280在1 8～2 0之间,蛋白、多糖、RNA等去除较彻底,适于AFLP分析。

The genomic DNA of young leaves of walnut were extracted with improved CTAB mothed.The extracted DNA was examined by ultraviolet absorption test and agarose gel electrophoresis. The results showed that CTAB (cetyltrimethyl ammonium bromide) buffer supplement with 3% PVP40(polyvinylpyrrolidone 40), 50 mmol/L β-mercaptoethanal and 10 mmol/L Na_2S_2O_5 was suitable for genomic DNA extraction in walnut.Then a method of DNA extracting in walnut could be found. The purity of the extracted DNA was very high with OD_(260)/OD_(280) value of 1.8～2.0.The protein, polyhexose and RNA were elminated completely. It could be used for AFLP analysis.