Transposition of native chromatin for personal epigenomics
DCFirst Claim
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1. A method for generating a sequencing library from a plurality of cells, comprising:
- a) lysing a plurality of cells to provide a plurality of cell nuclei, wherein the plurality of cell nuclei comprises chromatin;
b) contacting a cell nucleus of the plurality of cell nuclei with a transposase complex such that polynucleotides of the cell nucleus are tagmented at regions of open chromatin to produce a plurality of tagged fragments; and
c) performing one or more nucleic acid reactions on the tagged fragment to produce a sequencing library.
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Abstract
Provided herein is a method for analyzing polynucleotides such as genomic DNA. In certain embodiments, the method comprises: (a) treating chromatin isolated from a population of cells with an insertional enzyme complex to produce tagged fragments of genomic DNA; (b) sequencing a portion of the tagged fragments to produce a plurality of sequence reads; and (c) making an epigenetic map of a region of the genome of the cells by mapping information obtained from the sequence reads to the region. A kit for performing the method is also provided.
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29 Claims
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1. A method for generating a sequencing library from a plurality of cells, comprising:
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a) lysing a plurality of cells to provide a plurality of cell nuclei, wherein the plurality of cell nuclei comprises chromatin; b) contacting a cell nucleus of the plurality of cell nuclei with a transposase complex such that polynucleotides of the cell nucleus are tagmented at regions of open chromatin to produce a plurality of tagged fragments; and c) performing one or more nucleic acid reactions on the tagged fragment to produce a sequencing library. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. A method for generating a sequencing library from a plurality of cells, comprising:
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a) lysing a plurality of cells to isolate a plurality of cell nuclei, wherein the plurality of cell nuclei comprise chromatin; b) contacting a cell nucleus of the plurality of cell nuclei with a Tn5 transposase complex such that polynucleotides of the cell nucleus are tagmented at regions of open chromatin to produce a plurality of tagged fragments, wherein the Tn5 transposase complex comprises a first sequencing adapter sequence and a second sequencing adapter sequence, wherein the Tn5 transposase complex does not comprise an antibody specific to a protein that is part of chromatin, wherein a tagged fragment of the plurality of tagged fragment comprises i. a nucleotide sequence corresponding to a region of open chromatin, ii. the first sequencing adapter sequence, and iii. the second sequencing adapter sequence; and c) performing one or more nucleic acid reactions on the tagged fragment to produce a sequencing library. - View Dependent Claims (24, 25, 26, 27, 28, 29)
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Specification