北京油鸡MSAP毛细管电泳荧光检测技术的建立
收稿日期: 2013-09-29
修回日期: 2013-12-09
网络出版日期: 2014-05-25
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
Received date: 2013-09-29
Revised date: 2013-12-09
Online published: 2014-05-25
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。
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 . DOI: 10.3724/SP.J.1005.2014.0495
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.
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