CHEN Juan,JIN Wen-long,MO Yi-wei.Comparison on Genomic DNA Extracted from the Leaves of Chinese Chestnut by Two Methods[J].Northern Horticulture,2013,37(18):89-91.
两种方法提取板栗叶片DNA的效果比较
- Title:
- Comparison on Genomic DNA Extracted from the Leaves of Chinese Chestnut by Two Methods
- 文章编号:
- 1001-0009(2013)18-0089-03
- 分类号:
- S 603.6
- 文献标志码:
- A
- 摘要:
- 以采自湖北罗田地区的9个板栗品种为试材，研究比较了试剂盒法和改良CTAB法对板栗叶片DNA提取效果的影响。结果表明：用试剂盒提取的DNA出现部分降解的现象，A260/A280为1.11~1.49，DNA产率为24.50~59.25 μg/g，很难得到高纯度的DNA样品；而采用多次洗涤相结合，并用PVP和β-巯基乙醇联合除酚的改良CTAB法，能有效去除板栗叶片的多酚和多糖等物质，所获得的DNA完整，无降解现象，A260/A280为1.80~1.90，DNA产率为60.50~102.00 μg/g。表明改良的CTAB法获得的DNA纯度和产率较高，通过SRAP分子标记验证结果表明，DNA的纯度达到了板栗分子标记的试验要求。
- Abstract:
- New leaves of nine Castanea mollissima Blume varieties from Luotian country Hubei province were used as experimental materials and genomic DNA extraction methods of plant genomic DNA Kit and modified CTAB were used and compared.The results showed that the method of plant genomic DNA Kit was poor and was difficult to obtain clear bands by agarose gel electrophoresis.The ratio A260/A280of extracted DNA was 1.11~1.49 and the yield was 24.50~59.25μg/g.However,PVP and 2-mercaptoethanol were added to CTAB buffer in order to inhibit oxidation of polyphenol,and elution for many times were used to remove polysaccharide could be inhibited and most of polysaccharides in leaves could be removed efficiently.The ratio A260/A280of DNA extracted by modified CTAB ranged from 1.80~1.90,and the yield was 60.50~102.00 μg/g.The result of Castanea mollissima Blume molecular marker also showed that the modified CTAB method was suitable for extracting high qualified DNA from Castanea mollissima Blume.
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备注/Memo
第一作者简介：陈娟（1978-），女，博士，副教授，研究方向为植物生理与分子生物学。E-mail:breezy02008@163.com.
责任作者：莫亿伟（1971-），男，博士，副教授，研究方向为植物生理与分子生物学。E-mail:ywmo@163.com.
基金项目：经济林木种质改良与资源综合利用湖北省重点实验室开放基金资助项目（2011BLKF243；2013000503）。
收稿日期：2013-05-06