Methods of Genome Sequencing and Epigenetic Analysis

摘要

Novel methods of ChIP-seq are disclosed herein. These methods of ChIP-seq employ carrier DNA to prevent loss of DNA samples. The greater DNA yields achieved by this invention permit ChIP-seq of a small number of cells, permitting epigenetic analysis of primary cells of limited quantity.

主权项

1. A method of sequencing genomic DNA from a sample of cells, the method comprising:
a. Fragmenting chromatin in the sample of cells, b. Adding a carrier DNA to the fragmented chromatin of the sample of cells, wherein the carrier DNA is 5′ biotinylated DNA (“DNA1”), c. Precipitating the mixture of carrier DNA and fragmented chromatin, d. Annealing a blocking primer that is complementary to the DNA1, e. Amplifying the genomic DNA from the sample of cells, and f. Sequencing the amplified DNA; wherein the sample of cells comprise between 1 and 20,000 cells, and wherein the blocking primers prevent amplification of the DNA1.