公开(公告)号：US08440404B2

公开(公告)日：2013-05-14

申请号：US11071864

申请日：2005-03-03

申请人： Vladimir L. Makarov ,  Emmanuel Kamberov ,  Tong Sun ,  Jonathon H. Pinter ,  Brendan J. Tarrier ,  Eric E. Bruening ,  Takao Kurihara ,  Tim Tesmer ,  Joseph M'Mwirichia

发明人： Vladimir L. Makarov ,  Emmanuel Kamberov ,  Tong Sun ,  Jonathon H. Pinter ,  Brendan J. Tarrier ,  Eric E. Bruening ,  Takao Kurihara ,  Tim Tesmer ,  Joseph M'Mwirichia

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/6869 ,  C12N15/1072 ,  C12Q1/6827 ,  C12Q1/6855 ,  C12Q2525/301 ,  C12Q2525/191 ,  C12Q2521/331 ,  C12Q2523/125

摘要： The present invention regards a variety of methods and compositions for obtaining epigenetic information, such as DNA methylation patterns, through the preparation, amplification and analysis of Methylome libraries. In particular, the method employs preparation of a DNA molecule by digesting the DNA molecule with at least one methylation-sensitive restriction enzyme; incorporating a nucleic acid molecule into at least some of the digested DNA molecules by either (1) incorporating at least one primer from a plurality of primers that have a 5′ constant sequence and a 3′ variable sequence, wherein the primers are substantially non-self-complementary and substantially non-complementary to other primers in the plurality; or (2) incorporating an oligonucleotide having an inverted repeat and a loop under conditions wherein the oligonucleotide becomes blunt-end ligated to one strand of the digested DNA molecule, followed by polymerization from a 3′ hydroxyl group present in a nick in the oligonucleotide-linked molecule; and amplifying one or more of the DNA molecules.

摘要翻译： 本发明涉及通过Methylome文库的制备，扩增和分析来获得表观遗传信息的各种方法和组合物，例如DNA甲基化模式。 特别地，该方法通过用至少一种甲基化敏感性限制酶消化DNA分子来制备DNA分子; 通过（1）从具有5'恒定序列和3'可变序列的多个引物掺入至少一个引物，将核酸分子掺入到至少一些消化的DNA分子中，其中所述引物基本上不含 与多个中的其它引物互补且基本上不互补; 或者（2）在其中寡核苷酸变得钝端连接到消化的DNA分子的一条链上，然后从存在于寡聚核苷酸的切口中的3'羟基聚合的条件下，引入具有反向重复和环的寡核苷酸， 连接分子 并扩增一个或多个DNA分子。

公开(公告)号：US20050202490A1

公开(公告)日：2005-09-15

申请号：US11071864

申请日：2005-03-03

申请人： Vladimir Makarov ,  Emmanuel Kamberov ,  Tong Sun ,  Jonathon Pinter ,  Brendan Tarrier ,  Eric Bruening ,  Takao Kurihara ,  Tim Tesmer ,  Joseph M'Mwirichia

发明人： Vladimir Makarov ,  Emmanuel Kamberov ,  Tong Sun ,  Jonathon Pinter ,  Brendan Tarrier ,  Eric Bruening ,  Takao Kurihara ,  Tim Tesmer ,  Joseph M'Mwirichia

IPC分类号： C12N15/10 ,  C12P19/34 ,  C12Q1/68

CPC分类号： C12Q1/6869 ,  C12N15/1072 ,  C12Q1/6827 ,  C12Q1/6855 ,  C12Q2525/301 ,  C12Q2525/191 ,  C12Q2521/331 ,  C12Q2523/125

摘要： The present invention regards a variety of methods and compositions for obtaining epigenetic information, such as DNA methylation patterns, through the preparation, amplification and analysis of Methylome libraries. In several aspects of the present invention, there are methods based on methylation-dependent enrichment or depletion of genomic DNA isolated from cellular and cell-free sources. In additional embodiments, there are methods and compositions for single-step high throughput preparations of Methylome libraries.

摘要翻译： 本发明涉及通过Methylome文库的制备，扩增和分析来获得表观遗传信息的各种方法和组合物，例如DNA甲基化模式。 在本发明的几个方面，存在基于从细胞和无细胞来源分离的基因组DNA的甲基化依赖性富集或消耗的方法。 在另外的实施方案中，存在Methylome文库的单步高通量制剂的方法和组合物。