|Public on Jan 03, 2012
|Embryonic stem cells
|chip antibody: H3K27Ac
chip antibody manufacturer: Abcam
chip antibody catalog number: AB4729
cell line: V6.5 embryonic stem cells
|V6.5 (C57BL/6-129) murine ES cells were grown under typical ES conditions on irradiated mouse embryonic fibroblasts (MEFs). For location analysis, cells were grown for two passages off of MEFs, on gelatinized tissue-culture plates.
|Whole cell extracts were sonicated to solubilize the chromatin. The chromatin extracts containing DNA fragments with an average size of 500 bp were immunoprecipitated using different antibodies. Purified immunoprecipitated DNA were prepared for sequencing according to a modified version of the Solexa Genomic DNA protocol. Fragmented DNA was end repaired and subjected to 18 cycles of LM-PCR using oligos provided by Illumina. Amplified fragments between 150 and 300bp (representing shear fragments between 50 and 200nt in length and ~100bp of primer sequence) were isolated by agarose gel electrophoresis and purified.
|Illumina HiSeq 2000
|Chromatin IP against H3K27ac- ChIPSeq replicate 2
|Images analysis and base calling was done using the solexa pipeline.
For all samples reads were aligned to their indicated build using bowtie.
For all samples aligned sequences were extended 150bp upstream and 0bp downstream (with respect to read strand) and allocated into 25bp bins. Counts were normalized to reads per million, and bins with at least 1 read per million are shown.
|Dec 07, 2011
|Last update date
|May 15, 2019
|Richard A Young
|Whitehead Institute for Biomedical Research
|9 Cambridge Center
|Enhancer Decommissioning by LSD1 During Embryonic Stem Cell Differentiation (ChIP-seq)
|Enhancer Decommissioning by LSD1 During Embryonic Stem Cell Differentiation