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发明公开
CN105349528A 用于全基因组甲基化重亚硫酸氢盐测序的文库构建方法 有权 转让

用于全基因组甲基化重亚硫酸氢盐测序的文库构建方法
摘要：
本发明提供一种用于全基因组甲基化重亚硫酸氢盐测序的文库构建方法，其适用于中量级别(5～200ng)的起始样本量。该文库构建方法包括从5～200ng的基因组DNA得到片段化的基因组DNA；对上述片段化的基因组DNA进行重亚硫酸氢盐处理，得到单链的重亚硫酸氢盐处理产物；以上述单链的重亚硫酸氢盐处理产物为模板，使用含生物素标记的oligo1引物合成第一条链；使用核酸外切酶消化第一条链合成后体系中的单链DNA；使用链亲和素磁珠调取生物素标记的第一条链；以上述生物素标记的第一条链为模板，使用oligo2引物合成第二条链；以及，以所述第二条链作为模板进行PCR扩增，从而构建二代测序用DNA文库。
摘要（英）：

The invention provides a library construction method for whole genome methylation bisulfite sequencing. The method is suitable for a medium initial sample size (5-200 ng). The method comprises the steps of obtaining fragmented genome DNAs from 5-200 ng genome DNAs; conducting bisulfite treatment on the fragmented genome DNAs to obtain a single-stranded bisulfite treatment product; synthesizing a first chain by means of an oligo1 primer marked with biotin with the single-stranded bisulfite treatment product as the template; digesting single-stranded DNAs in a system after the first chain is synthesized with exonuclease; obtaining the first chain marked with biotin by means of streptavidin magnetic beads; synthesizing a second chain by means of an oligo2 primer with the first chain marked with biotin as the template; conducting PCR amplification with the second chain as the template, so that a DNA library for next-generation sequencing is constructed.

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