2025-06-19
ENA first public
ERC000011
ENA-CHECKLIST
University of Toronto, Hospital for Sick Children
INSDC center name
public
INSDC status
ERS22185479
SRA accession
E-MTAB-14612:Jun_2_cow
Submitter Id
ArrayExpress
broker name
Bovine Aortic Endothelial Cells
cell_line
not collected
collection date
domestic cattle
common name
Protocols: Chromatin immunoprecipitation experiments were conducted as previously described in (Schmidt et al. 2009). Antibodies used for ChIP were mouse anti-H3K27ac (Millipore, 05-1334 monoclonal)and rabbit anti-Jun (Santa Cruz Biotechnology, sc1694 polyclonal). EC were grown in Endothelial Cell Medium (ScienCell, catalogue #1001) ChIP DNA was prepared for Illumina sequencing by blunt-end repair, dA-tailing, and ligation of Illumina adaptors using a NEBNext DNA library preparation kit (New England Biolabs, catalogue #E6040L). Total ChIP DNA (approximately 200-500ng) and 220ng of DNA input (WCE) was end repaired for 30 minutes at room temperature, and then purified using column purification with either DNA Clean and Concentrator (Zymogen, catalogue #D4014) or PCR purification columns (Qiagen, catalogue #28106) as recommended by manufacturer’s protocol. Blunt-end repaired DNA was dA-tailed for 40 minutes at 37° C, then column purified. dA-tailed DNA was ligated to Illumina adaptors (final concentration 6.67 nM). USER enzyme was used to cleave the uracil hairpin of the Illumina adaptor, and adaptor-ligated DNA was column purified. The library was PCR amplified for 16-18 cycles using a universal primer and a barcoded primer (New England Biolabs, catalogue #E7335L). PCR-amplified DNA was purified using PCR column purification and eluted in 20 uL of elution buffer for preparation of gel extraction, or 30 uL of TE for preparation of Pippin Prep size selection. Libraries were size selected from 200-350bp using a 2% agarose gel or a dye-free automated size selection cassette from Pippin Prep (Sage Sciences, catalogue #CDF2010).
description
not collected
geographic location (country and/or sea)
not applicable
isolate
Bos taurus
organism
Bos taurus
organism
Bos taurus
scientific_name
unknown
sex
Jun_2_cow
title