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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (22): 236-241.doi: 10.11924/j.issn.1000-6850.2011-3766

所属专题： 生物技术

• 林学 园艺 园林 • 上一篇    下一篇

款冬花DNA提取及ISSR体系优化

贺润丽   

  • 收稿日期:2011-12-13 修回日期:2012-02-24 出版日期:2012-08-05 发布日期:2012-08-05

Genomic DNA Isolation and Optimization for ISSR Reaction System of Tussilago farfara L.

  • Received:2011-12-13 Revised:2012-02-24 Online:2012-08-05 Published:2012-08-05

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摘要：

摘 要：为了从分子水平对款冬种质资源进行鉴定和遗传多样性分析，分别采用SDS法、CTAB法、改良SDS法和改良CTAB法提取款冬叶片基因组DNA，比较不同方法的提取效果，并用改良CTAB法提取的DNA进行ISSR-PCR扩增体系摸索，建立优化的ISSR-PCR反应体系。结果显示：改良SDS法和改良CTAB法都可以提取出高质量的DNA，最优的ISSR-PCR反应条件为: 20μl反应体系中含DNA模板约20ng，Taq 酶1.0 U，Mg2+ 2.00 mmol/L，dNTPs 0.20 mmol/L，引物0.6 μmol/L，1×PCR缓冲液。

关键词: 产量, 产量

Abstract:

To investigate the genetic diversity and identification of germplasm resources, the genomic DNA was extracted from the leaves of Tussilago farfara L. by using the SDS, CTAB, modified SDS and modified CTAB methods. The extraction effects of different methods were compared. The subsequent ISSR-PCR condition was explored, using the DNA harvested from modified CTAB methods. The results showed that the modified SDS and CTAB methods could effectively obtain high quality DNA, and the most suitable mixture was as follows: total 20μl reaction volume containing template DNA 20ng, 1.0 UTaq DNA polymerase, 2.00 mmol/L Mg2+, 0.20 mmol/L dNTPs, 0.6 μmol/L pimer, 1×PCR buffer.