GFP_Trap_2
Mus musculus
Protocols: ChIP samples were pre-cleared and then incubated with GFP-Trap MA at 4 degrees overnight. Precipitates were then washed to remove non-specifically bound proteins and DNA 5 x 10^7 CD4 T cells from GFP-Egr2 mice were stimulated with anti-CD3 and anti-CD28 for 24 before harvesting Cells were crosslinked with 1% Formaldehyde for 10 min at room temperature and the reaction was then quenched with 125mM Glycine Chromatin was sheared by sonication with a Bioruptor Pico (Diagenode) Crosslinks were reversed and protein degraded by heating to 65C in the presence of Proteinase K. DNA was then purified by phenol chloroform extraction and ethanol precipitation The libraries were generated from the DNA using the NEBNext Ultra II DNA library Prep kit (New England Biolabs) according to the manufacturer's instructions