|本期目录/Table of Contents|

[1]蔡志翔,沈志军,严娟,等.桃甲基化敏感扩增多态性(MSAP)技术体系的建立[J].江苏农业科学,2016,44(11):43-45.
 Cai Zhixiang,et al.Establishment of peach methylation sensitive amplified polymorphism detection system[J].Jiangsu Agricultural Sciences,2016,44(11):43-45.
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桃甲基化敏感扩增多态性(MSAP)技术体系的建立(PDF)
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《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:
第44卷
期数:
2016年11期
页码:
43-45
栏目:
生物技术
出版日期:
2016-11-25

文章信息/Info

Title:
Establishment of peach methylation sensitive amplified polymorphism detection system
作者:
蔡志翔 沈志军 严娟 马瑞娟 俞明亮
江苏省农业科学院园艺研究所/江苏省高效园艺作物遗传改良重点实验室,江苏南京 210014
Author(s):
Cai Zhixianget al
关键词:
DNA甲基化MSAP体系优化
Keywords:
-
分类号:
S188;S662.101
DOI:
-
文献标志码:
A
摘要:
为建立适合桃的甲基化敏感扩增多态性(methylation sensitive amplification polymorphism,简称MSAP)反应体系,以桃PCM-1R、PCM-1G为材料,对MSAP技术中的关键因素进行优化。结果表明,500 ng基因组DNA用EcoRⅠ、HapⅡ或MspⅠ各10 U(0.5 μL,2 000 U/mL),37 ℃保温12 h,即可酶切完全;25 μL选择性扩增体系中,含有2 μL 10倍稀释的预扩增产物、各1 μL上下游引物、2.5 μL 10×Taq buffer、2.5 μL dNTP mix(各0.2 mmol/L)、2.5 μL 25 mmol/L MgCl2、0.25 μL Taq DNA聚合酶。在该体系下选用256对引物对桃叶片进行甲基化模式分析,经筛选获得23对扩增条带清晰、重复性好的引物,PCM-1R、PCM-1G总甲基化率分别为28.0%、25.7%。
Abstract:
-

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2015-10-28
基金项目:国家自然科学基金(编号:31101517);江苏省农业科技自主创新资金[编号:CX(12)5034]。
作者简介:蔡志翔(1979—),男,江苏南京人,硕士,助理研究员,主要从事桃种质资源收集、保存、评价工作。Tel:(025)84390855;E-mail:czx1y05@163.com。
更新日期/Last Update: 2016-11-25