Patent ID: 11965890
Assignee: THE UNIVERSITY OF CHICAGO
Field: Measurement (Instruments)
Classification: CPC G  C | IPC C  G

Claim 0:
1. A method of determining a density of a first epitope of a core histone at a genomic locus in chromatin of a cell, the method comprising:
preparing a library of native nucleosomes from the chromatin, wherein the library comprises nucleosomes, each comprising the core histone and a nucleosome nucleotide sequence indicative of its genomic locus of origin;
adding a standard to the library to create a doped library; wherein the standard comprises a reconstituted nucleosome comprising (i) a standard histone or standard histone fragment having the first epitope and (ii) a standard molecule comprising a barcode molecule, wherein the standard histone or standard histone fragment and the standard molecule form a stable protein-DNA association;
determining an amount of the core histone at the genomic locus in the doped library;
determining an amount of the standard in the doped library;
adding a first affinity reagent to the doped library to capture an amount of native nucleosomes and reconstituted nucleosomes comprising the epitope;
determining a relative genomic abundance for the first epitope at a genomic locus based on the amount of the captured standard comprising the epitope and the amount of the core histone at the genomic locus in the doped library;
determining a standard capture efficiency for the epitope based on the amount of captured standard compared to the amount of standard added to the doped library;
determining the relative genomic abundance of the first epitope of the core histone at the genomic locus based on a first epitope abundance for the core histone and the standard capture efficiency.