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US 8,076,070 B2Granted
Genome-wide chromosome conformation capture
AssigneeUniversity of Southern California
InventorsLin Chen, Reza Kalhor
Life of this patent
Estimating the term…
Abstract
The invention relates to the use of GCC (Genome Conformation Capture) technology in determining the three dimensional arrangement of an entire genome.
Claims15
A method of determining the three-dimensional arrangement of an entire genome in a cell, comprising:
- (a) contacting a cell with a cross-linking reagent to cross-link the chromatin and the proteins in the cell;
- (b) lysing the cell;
- (c) digesting the chromatin with a restriction enzyme to generate DNA fragments with free ends, wherein the DNA fragments are cross-linked to the proteins;
- (d) ligating DNA linkers to all free DNA ends generated by the restriction enzyme;
- (e) annealing the ends of the DNA linkers that are not ligated to the free DNA ends generated by the restriction enzyme;
- (f) translating the nicks between the annealed ends of the DNA linkers;
- (g) detaching the proteins cross-linked to the DNA fragments;
- (h) extracting DNA;
- (i) subjecting the extracted DNA to two rounds of parallel sequencing;
- and
- (j) determining the three-dimensional arrangement of the entire genome in the nucleus of the cell.
8 more elements
The method of clam 1, wherein the cross-linking reagent is formaldehyde.
Dependencies are inferred from the claim text.