北京油鸡MSAP毛细管电泳荧光检测技术的建立

doi:10.3724/SP.J.1005.2014.0495

遗传 ›› 2014, Vol. 36 ›› Issue (5): 495-502.doi: 10.3724/SP.J.1005.2014.0495

• 技术与方法 • 上一篇

北京油鸡MSAP毛细管电泳荧光检测技术的建立

李金龙¹, 唐韶青², 邹智元³, 王海潮¹, 徐青¹

1. 北京交通大学生命科学与生物工程研究院, 北京 100044;
2. 北京市畜牧总站, 北京 100029;
3. 北京交通大学软件学院, 北京 100044

收稿日期:2013-09-29 修回日期:2013-12-09 出版日期:2014-05-20 发布日期:2014-05-25
通讯作者: 徐青, 博士, 副教授, 研究方向：分子遗传学。E-mail: qingxu@bjtu.edu.cn E-mail:qingxu@bjtu.edu.cn
作者简介:李金龙, 硕士研究生, 专业方向：分子遗传学。E-mail: lijinlong880827@163.com
基金资助:
国家自然科学基金青年基金项目(编号：31101711), 北京市自然科学基金项目(编号：6112018)和中央高校基本科研业务费专项基金项目(编号：2012JBZ003)资助

Establishment of fluorescence labeling and capillary electrophoresis in MSAP for Beijing You chicken

Jinlong Li¹, Shaoqing Tang², Zhiyuan Zou³, Haichao Wang¹, Qing Xu¹

1. Institute of Life Science and Biotechnology, Beijing Jiaotong University, Beijing 100044, China;
2. Beijing Animal Husbandry Station, Beijing 100029, China;
3. Institute of Software, Beijing Jiaotong University, Beijing 100044, China

Received:2013-09-29 Revised:2013-12-09 Online:2014-05-20 Published:2014-05-25

摘要/Abstract

摘要：

采用毛细管电泳荧光检测技术, 对北京油鸡肌肉组织基因组进行甲基化敏感扩增片段多态性(Methylation sensitive amplified polymorphism, MSAP)检测。通过对基因组DNA用量、预扩产物稀释倍数、选择性引物浓度、Mg2+浓度、dNTPs浓度和电泳内标量等6个主要参数进行分析和优化, 建立了适合北京油鸡基因组DNA甲基化分析的MSAP毛细管电泳荧光检测技术。重复实验表明, 建立的毛细管电泳荧光标记的MSAP检测技术能够自动地、高通量地分析北京油鸡基因组的DNA甲基化状态, 也适用于其他动植物基因组的DNA甲基化研究。

关键词: MSAP, 毛细管电泳, 荧光标记, 北京油鸡

Abstract:

The DNA methylated states in muscle tissues from Beijing You chicken were analyzed using methylation sensitive amplified polymorphism (MSAP) combined with fluorescence labeling and capillary electrophoresis technology. We optimized the MSAP experimental condition including the genomic DNA concentration, dilution of pre-amplification prod-uct, the internal standard content and the concentrations of selective primers, Mg2+ and dNTPs. Repeated experiments showed that this method can automatically detect the global DNA methylation states in Beijing You chicken, and can be extended to other animals or plants with complex genomes and rich methylation polymorphism.

Key words: methylation sensitive amplified polymorphism (MSAP), capillary electrophoresis, fluorescence labeling, Beijing You chicken

李金龙, 唐韶青, 邹智元, 王海潮, 徐青. 北京油鸡MSAP毛细管电泳荧光检测技术的建立[J]. 遗传, 2014, 36(5): 495-502.

Jinlong Li, Shaoqing Tang, Zhiyuan Zou, Haichao Wang, Qing Xu. Establishment of fluorescence labeling and capillary electrophoresis in MSAP for Beijing You chicken[J]. HEREDITAS(Beijing), 2014, 36(5): 495-502.

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北京油鸡MSAP毛细管电泳荧光检测技术的建立

Establishment of fluorescence labeling and capillary electrophoresis in MSAP for Beijing You chicken

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