Rubicon Genomics, Inc.

Jonathan H Pinter

Vladimir L Makarov

Emmanuel Kamberov

Brendan J Tarrier

Eric Bruening

Takao Kurihara

Tong Sun

Tim Tesmer

Joseph M'Mwirichia

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&#x2032; 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&#x2032; hydroxyl group present in a nick in the oligonucleotide-linked molecule; and amplifying one or more of the DNA molecules.

Methods and compositions for generating and amplifying DNA libraries for sensitive detection and analysis of DNA methylation

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Ypsilanti, MI, United States of America

Jonathan H Pinter