GSM4972504: YTHDF2 kockdown(KD) cells for STAT5a IP; Homo sapiens; ChIP-Seq
add 37% formaldehyde to the culture medium with at least 107 cells(final concentration of formaldehyde is 1%), shake on a shaker for 10 minutes; stop formaldehyde fixation: add glycine solution to a final concentration of 0.125mol/L, pipette and mix;Wash the cells with PBS, scrape the cells, harvest the cells, and discard the supernatant as much as possible.Combine the antibody with protein A+G magnetic beads (Magna ChIP™ 16-663);Add lysate and protease inhibitor to the cells, and sonicate in ice water bath; take 20ul as Input sample and store at 4℃;Add the sample to the protein A+G magnetic beads bound to the antibody and mix overnight; wash the magnetic column to elute the fragments bound to the protein A+G magnetic beads, and Input samples were added to ProteinaseK together and de-crosslinked at 65℃ overnight; DNA was extracted with phenol chloroform and digested with RNase. Repair DNA fragment ends, add A base at 3'end, connect sequencing adapter; PCR amplification and fragment size selection of DNA productsm;After the library is qualified, it is sequenced through the Illumina HiSeq/NextSeq platform.
GEO Accession
GSM4972504