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公开(公告)号：US09920363B2
公开(公告)日：2018-03-20
申请号：US14358674
申请日：2012-11-15
发明人： Fei Gao , Junwen Wang , Tong Wang , Hui Jiang , Jinghua Wu , Honglong Wu
CPC分类号： C12Q1/6869 , C12N15/1093 , C12Q1/6806 , C12Q1/6876 , C40B40/06 , C40B40/08 , C40B50/06 , C12Q2523/125 , C12Q2525/117 , C12Q2525/191 , C12Q2535/122
摘要： The present invention provides a method for constructing a high-throughput sequencing library, which comprises: fragmenting genomic DNA; end-repairing the DNA fragments; adding a base A to the 3′ end of the end-repaired DNA fragments; connecting the DNA fragments having cohesive end A with a methylated adapter; carrying out hybrid capture on the connection products by using specific probes to obtain object fragments; treating the object fragments with bisulfite, to convert non-methylated cytosines to uracils; PCR amplifying the converted object fragments; and separating and purifying the amplification products, wherein the amplification products constitute the high-throughput sequencing library. The present invention also provides a method and an apparatus for identifying methylation information in specified genome regions of a sample.
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2.发明申请公开(公告)号：US20140329697A1
公开(公告)日：2014-11-06
申请号：US14358674
申请日：2012-11-15
发明人： Fei Gao , Junwen Wang , Tong Wang , Hui Jiang , Jinghua Wu , Honglong Wu
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6869 , C12N15/1093 , C12Q1/6806 , C12Q1/6876 , C40B40/06 , C40B40/08 , C40B50/06 , C12Q2523/125 , C12Q2525/117 , C12Q2525/191 , C12Q2535/122
摘要： The present invention provides a method for constructing a high-throughput sequencing library, which comprises: fragmenting genomic DNA; end-repairing the DNA fragments; adding a base A to the 3′ end of the end-repaired DNA fragments; connecting the DNA fragments having cohesive end A with a methylated adapter; carrying out hybrid capture on the connection products by using specific probes to obtain object fragments; treating the object fragments with bisulfite, to convert non-methylated cytosines to uracils; PCR amplifying the converted object fragments; and separating and purifying the amplification products, wherein the amplification products constitute the high-throughput sequencing library. The present invention also provides a method and an apparatus for identifying methylation information in specified genome regions of a sample.
摘要翻译： 本发明提供了构建高通量测序文库的方法，其包括：基因组DNA的片段化; 最终修复DNA片段; 在最终修复的DNA片段的3'端加入碱基A; 将具有粘性末端A的DNA片段与甲基化衔接子连接; 通过使用特定的探针对连接产品进行混合捕获以获得对象片段; 用亚硫酸氢盐处理物体碎片，将非甲基化的胞嘧啶转化成尿嘧啶; PCR扩增转化的对象片段; 并分离和纯化扩增产物，其中扩增产物构成高通量测序文库。 本发明还提供了用于鉴定样品的指定基因组区域中的甲基化信息的方法和装置。
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