烤后烟叶基因组DNA提取条件优化

马林; 王广超; 罗昭标; 寇晓腾; 张文龙

doi:10.3969/j.issn.2095-476X.2013.02.014

March 2013

Article Contents

MA Lin, WANG Guang-chao, LUO Zhao-biao, et al. The optimization of conditions for the extraction of genomic DNA in flue-cured tobacco[J]. Journal of Light Industry, 2013, 28(2): 59-63. doi: 10.3969/j.issn.2095-476X.2013.02.014

Citation: MA Lin, WANG Guang-chao, LUO Zhao-biao, et al. The optimization of conditions for the extraction of genomic DNA in flue-cured tobacco[J]. Journal of Light Industry, 2013, 28(2): 59-63. doi: 10.3969/j.issn.2095-476X.2013.02.014 shu

The optimization of conditions for the extraction of genomic DNA in flue-cured tobacco

College of Tobacco Science and Engineering, Zhengzhou University of Light Industry, Zhengzhou 450001, China;
Tobacco Management Department, Zixi Branch Fuzhou Tobacco Company, Fuzhou 335300, China

Received Date: 2013-01-02
Available Online: 2013-03-15

Abstract

The experimental method which improves the CTAB for extracting the genomic DNA in flue-cured tobacco was presented in order to stabilize the extraction of the genomic DNA. Each of the four influencing factors in the CATB, namely, the usage amount of precipitate buffer solution, pyrolysis time,number of processing the Tris phenol and the processing time for RNase were optimized respectively. The results showed that after using CATB precipitate buffer solution with the volume 1.5 times larger than that of the DNA solution with pyrolysis time for 40 min, one-time extraction of the Tris phenol and 10 min processing the Rnase (10 mg/mL,1 μL), a clear main band of the genomic DNA in the flue-cured tobacco could be obtained. The extracted genomic DNA OD₂₆₀/OD₂₈₀ was between 1.7 and 1.9. OD₂₆₀/OD₂₃₀ was greater than 2.0. In this case, with 35 ng genomic DNA, clear RAPD and SCAR maps could be obtained. The extracted genomic DNA can be applied to the molecular biological experiments.
- flue-cured tobacco,
- genomic DNA,
- extraction of DNA,
- PCR
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The optimization of conditions for the extraction of genomic DNA in flue-cured tobacco

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通讯作者: 陈斌, bchen63@163.com