适于AFLP分析用的桃韧皮部DNA提取方法

摘要

[Objective] The research aimed to study the peach phloem DNA extraction method which was suitable for AFLP analysis,which laid the foundation for the peach genome research when the spire and other young materials couldn't be obtained in winter. [ Method] The phloem of eight kinds of wild peach dormant wood in one year old in winter was the test material. The genome total DNA was extracted by u-sing the improved CTAB method,and the extraction effect was compared with that in the corresponding spire DNA. Moreover,AFLP verified a-nalysis was carried out. [Result] Except PNA yield which was extracted from the wild peach phloem was lower than that in the spire,DNA purity and integrity were good. OD260/OD280 was during 1.8-2. 0,and there was no degradation phenomenon. RNA removal was clean,and it could be completely digested by the restriction endonuclease. AFLP analysis band was clear,and the polymorphism was good. [ Conclusion] The wild peach phloem genome total DNA which was extracted by the method was entirely suitable for AFLP analysis.%[目的]研究适于AFLP分析用的桃韧皮部DNA提取方法,为冬季无法获得幼叶和其他幼嫩材料时从事桃基因组研究打下了基础.[方法]以8种野生桃冬季1年生休眠枝的韧皮部为试验材料,采用改进的CTAB法提取基因组总DNA,与其相应幼叶DNA的提取效果进行了比较分析,并进行了AFLP验证分析.[结果]从野生桃韧皮部提取的DNA除产量低于幼叶外,DNA纯度和完整性都较好,OD(260)/OD280均为1.8～2.0,无降解现象,RNA去除干净,能被限制性内切酶完全消化,其AFLP分析条带清晰,多态性好.[结论]该法提取的野生桃韧皮部基因组总DNA完全适于AFLP分析.