摘要
[目的]筛选出玉米胚乳DNA提取的最佳方法。[方法]采用改进的CTAB提取液,以高温和常温2种方法提取玉米胚乳基因组DNA,用紫外分光光度计、0.8%水平的琼脂糖凝胶电泳及PCR检测法比较了2种方法的提取效果。[结果]高温法所提玉米胚乳DNA浓度和纯度均达到分子生物学要求,而常温法所提DNA的纯度未达到分子生物学的要求。电泳检测表明,高温法提取的DNA均未降解,DNA条带整齐度较好,而用常温法提取的DNA都出现了降解。PCR检测结果表明,用高温法提取的DNA作模板的PCR所得条带清晰,明亮,而用常温法提取的DNA作模板的PCR条带不清晰。最佳聚乙烯聚吡咯烷酮（PVPP）的浓度确定为玉米籽粒重的6%～10%。[结论]高温法更适宜用作玉米胚乳DNA提取。
[ Objective] The aim of the study was to screen out the optimum method of extracting maize endosperm DNA. [ Method] The improved CrAB extract was used to extract the total genome DNA from maize endosperm with high temperature and normal temperature method. And their extracting effect were compared by the test methods of ultraviolet spectrophotometer, agaroseGel electrophoresis at 0.8% level and PCR. [ Result] The concn, and purity of the maize endosperm DNA extracted by high temperature method all accorded with the demand of the optimal molecular biology, while the purity of DNA extracted by normal temperature method didn＇ t accorded with the demand of the optimal molecular. The electropboresis test. showed that there was no degradation in DNA extracted by high temperature method and DNA brands had better uniformity, while the degradation appeared in DNA extracted by normal temperature method. PCR test showed that taking DNA extracted by high temperature method as the templet could get clear PCR bands and taking DNA extracted by normal temperature method as the templet couldn＇ t get clear PCR bands, The optimum concn, of PVPP was ascertained as 6% - 10% of maize grain weight. [ Conclusion] The high temperature method was mere suitable for extracting maize endosperm DNA.
出处
《安徽农业科学》
CAS
北大核心
2008年第22期9408-9409,共2页
Journal of Anhui Agricultural Sciences
基金
北京市科技新星资助项目(2007B033)
