[1]杨晓婷,黄建,张春梅,等. 枣叶绿体基因组DNA提取方法研究[J].西北林学院学报,2015,30(02):105-110.[doi:doi:10.3969/j.issn.1001-7461.2015.02.18]
 YANG Xiao-ting,HUANG Jian,ZHANG Chun-mei,et al. An Optimized Chloroplast DNA Extraction Protocol for Chinese Jujube[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2015,30(02):105-110.[doi:doi:10.3969/j.issn.1001-7461.2015.02.18]
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 枣叶绿体基因组DNA提取方法研究()
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《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]

卷:
第30卷
期数:
2015年02期
页码:
105-110
栏目:
出版日期:
2015-03-30

文章信息/Info

Title:
 An Optimized Chloroplast DNA Extraction Protocol for Chinese Jujube
文章编号:
1001-7461(2015)02-0105-06
作者:
 杨晓婷黄建张春梅李新岗*
 (西北农林科技大学 林学院,国家林业局枣工程技术研究中心,陕西 杨陵 712100)
Author(s):
 YANG Xiao-ting HUANG Jian ZHANG Chun-mei LI Xin-gang*
 (College of Forestry, Centre of Jujube Engineering and Technology of State Forestry Administration, Northwest A&F University, Yangling, Shaanxi 712100 , China)
关键词:
 叶绿体基因组cpDNA提取改良高盐-低pH法
Keywords:
Ziziphus jujuba chloroplast genome cpDNA extraction modified high-salt low-pH method
分类号:
S665.1
DOI:
doi:10.3969/j.issn.1001-7461.2015.02.18
文献标志码:
A
摘要:
 提取高质量的叶绿体基因组DNA是叶绿体基因组测序分析的重要环节。研究以冬枣、晋枣成熟叶片为材料,采用高盐-低pH法和改良的高盐-低pH法比较研究枣叶绿体分离和叶绿体DNA(cpDNA)提取。结果表明,高盐-低pH法提取的cpDNA,其OD260nm/OD280nm值均为1.28,质量低,不能满足后续测序要求;改良高盐-低pH法提取的cpDNA产量明显得到提高,且cpDNA结构完整、质量高、纯度好,OD260nm/OD280nm值介于1.8~2.0,无降解现象,RNA消化完全,能满足后续测序要求。
Abstract:
 Choloroplast genomes supply valuable genetic information for evolutionary and functional studies in plants; However, extraction of high quality chloroplast DNA (cpDNA) is a critical step before chloroplast genomes sequencing. Mature leaves collected from two cultivars (Dongzao and Jinzao) were used for chloroplast and cpDNA isolation in present study. High-salt low-pH method and a modified high-salt low-pH method were utilized for cpDNA isolation. The results showed: the value of OD260nm/OD280nm of cpDNA isolated by the original method was 1.28, therefore, it can’t meet the requirements of subsequent chloroplast genomes sequencing. However, the yields of cpDNA extraction by the modified method were dramatically higher than that by the routine method, and the extracted cpDNA of mature leaves by this improved method was integral, high-quality and pure. In addition, the value of OD260nm/OD280nm was 1.8 to 2.0, degradation was nearly free, and the RNA was eliminated completely. Thus, the quality of cpDNA extracted by this modified high-salt low-pH method can meet the requirements of subsequent chloroplast genomes sequencing.

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备注/Memo

备注/Memo:
 收稿日期:2014-06-11修回日期:2014-06-17
基金项目:国家林业局公益性行业专项(201304110);陕西省自然科学基金(2013JQ3001)。
作者简介:杨晓婷,女,在读硕士生,研究方向:枣树叶绿体基因组。E-mail: yangxiaoting@nwsuaf.edu.cn
*通信作者:李新岗,男,教授,博士生导师,研究方向:红枣产业技术。E-mail: xingangle@nwsuaf.edu.cn
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