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中国农学通报 ›› 2012, Vol. 28 ›› Issue (36): 46-49.

所属专题: 生物技术 小麦

• 农学 农业基础科学 • 上一篇    下一篇

一种快速提取小麦基因组DNA的改良CTAB方法

张晓祥 王玲 寿路路   

  • 收稿日期:2012-08-03 修回日期:2012-09-11 出版日期:2012-12-25 发布日期:2012-12-25
  • 基金资助:

    抗病虫转基因小麦新品种培育;抗逆转基因小麦新品种培育

A Rapid Modified CTAB Method of Extracting Genomic DNA from Wheat Leaf

  • Received:2012-08-03 Revised:2012-09-11 Online:2012-12-25 Published:2012-12-25

摘要:

旨在寻求一种微量、快速提取小麦DNA的方法。以小麦幼叶为材料,用改良CTAB法、改良SDS法、沸水浴法提取小麦叶片总DNA,通过琼脂糖凝胶电泳法、紫外吸收法和PCR检测DNA完整性和纯度。改良CTAB法提取小麦基因组DNA质量和纯度高、无降解,每人每天可以轻松提取200个DNA样品,对转基因植株的PCR检测显示扩增目标条带清晰一致,无假阳性,试验结果理想。改良SDS法所提DNA纯度和浓度虽略不如改良CTAB法,但仍然符合实验要求。沸水浴法提取量少且杂质多,PCR无有效扩增,结果不理想。改良CTAB法提供了一种简便、快速微量小麦DNA提取方法,适用于PCR和其他分子生物学研究。

关键词: 质量, 质量

Abstract:

The aim was to seek for a rapid DNA miniprep extraction method from wheat leaves. The wheat leaves were used as experimental materials, total DNA was extracted by using methods including modified CTAB, modified SDS and Boiling. Integrality and purity of nucleic acids were detected with agarose gel electrophoresis, ultraviolet absorption and PCR. DNA quality and purity extracted by modified CTAB method were high and had no degradation phenomenon. Two hundred DNA samples could be extracted each workday by per capita using this method. PCR detection of wheat transgenic plants showed that amplified bands of target gene were clear, without false-positive, and the test results were satisfactory. The DNA purity and concentration extracted by modified SDS method were not as good as modified CTAB method, but it also met the DNA requirements of major molecular research. The DNA quantity extracted by modified Boiling method was small and had a lot of impurities in it, PCR detection of this DNA showed no amplified band, the test results were unsatisfactory. Modified CTAB method provided a simple, rapid and miniprep method for extracting DNA from wheat, and was suitable for PCR amplification and other molecular biology research.

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