鹿茸干细胞基因组DNA甲基化的检测方法

以鹿茸干细胞为研究对象,分别采用甲基化敏感性扩增多态性(MSAP)和荧光标记甲基化敏感性扩增多态性(F-MSAP)方法对鹿茸干细胞进行全基因组DNA甲基化检测。结果表明:MSAP方法共检测到387个位点,F-MSAP方法共检测出1 524个位点。2种方法所得结果中均发现Ⅰ型条带数最多,Ⅲ型条带数最少。与MSAP方法比较,F-MSAP方法在数据分析和实际操作过程中具有安全、高效、高通量、自动化等特点,更适用于检测鹿茸干细胞基因组DNA甲基化。

Comparative Study on Methods of Genome-wide DNA Methylation Assay in Antler Stem Cells

The methylation-sensitive amplification polymorphism ( MSAP ) and the fluorescence-la-beled methylation-sensitive amplification polymorphism ( F-MSAP ) are both modified AFLP ( ampli-fied fragment length polymorphism) techniques to investigate cytosine methylation in genomes. In F-MSAP system,selective amplification is fluorescently labeled and selective products could be detec-ted by DNA sequencer. In the present study, MSAP and F-MSAP methods were used to analyze DNA methylation in antler stem cells. In MSAP and F-MSAP systems, a total of 387 and 1 524 frag-ments were detected, respectively, and type I fragments are the most frequent and typeⅢfragments are the least frequent. The results show that compared to MSAP method, F-MSAP method had the main advantages of safety, high efficiency, high sensitivity and automation in the process of data a-nalysis and operation. Overall, the present study reveals that the F-MSAP method was more suitable than MSAP method for detecting cytosine methylation in antler stem cells as well as deer.