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4种水稻基因组DNA微量提取方法对ISSR-PCR试验的影响

     

摘要

[Objective] The good and simple method of DNA extraction, which was suitable for the ISSR amplification experiment, was ex-plored. [Method] The purity and concentration of the genomic DNA, extracted-with two methods of SDS and CTAB, and two types of kits (DP305 and DP320), were measured with the nucleic acid proteb detector and agarose gel electrophoresis, and then, the efficiency of the IS-SR-PCR application based on them as template was compared. [ Result] It was found that the concentration and purity of DNA extracted with SDS and CTAB were not fine, which could be amplified the formation of ISSR-PCR bands but there was a poor reliability. The application of Kit DP305 and DP320 could result in a fine extraction effect on both DNA concentration and purity, among which, the better result could be produced from Kit DP320 with stable reliability. [Conclusion] The DNA extracted with the kit DP320 was most suitable fot the rice ISSR ex-periment.%[目的]找出适用于ISSR试验的效果好、操作简便的水稻DNA提取方法.[方法]采用高盐十二烷基硫酸钠(SDS)法、十六烷基三甲基溴化铵( CTAB)法及2种型号试剂盒(DP305、DP320)法提取基因组DNA,利用核酸蛋白检测仪及琼脂糖电泳法测定其纯度与浓度,并比较其作为模板用于ISSR-PCR的效果.[结果]发现SDS法与CTAB法提取的DNA浓度与纯度均不高,在1SSR-PCR中虽能扩增出条带,但结果不能重复;试剂盒DP305与DP320提取的DNA浓度与纯度较好,其中试剂盒DP320效果更优,PCR结果能稳定重复.[结论]试剂盒DP320提取DNA的方法是最适合水稻ISSR试验的DNA提取方法.

著录项

  • 来源
    《安徽农业科学》|2011年第24期|14540-14541|共2页
  • 作者

    张静;

  • 作者单位

    江汉大学生命科学学院,湖北武汉430056;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 S188+.1;
  • 关键词

    DNA; ISSR-PCR; 水稻;

  • 入库时间 2022-08-17 11:50:11

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