GSM3854231: cKO_3 ATAC; Mus musculus; ATAC-seq
For ChIP-seq, FACS-sorted cells were crosslinked with 1% formaldehyde at room temperature with gentle agitation for 10 minutes. The crosslink was terminated by Glycine at a final concentration of 0.125 M. Cells were then washed three times with ice-cold PBS and stored at -80°C until a sufficient number of cells had been obtained. ChIP was performed following standard ChIP protocols with the following major modifications: 10E6 cells were resuspended in 200 ml lysis buffer and sonicated to obtain DNA fragments from 150 to 500 bp in size. Each ChIP reaction was performed with 10E6 cells and 5 g of antibody. For ATAC-seq, transposed genomic DNA was generated with Illumina Nextera DNA Library Prep Kit and purified with Qiagen MinElute Reaction Cleanup Kit. ChIP-seq libraries were generated with Nugen Ovation Ultralow System v2. ATAC-seq libraries were constructed with NEBNext High-Fidelity 2x PCR Master Mix in the presence of specific barcoding primers.
GEO Accession
GSM3854231