GSM8475016: Kidney_C24V1V2_pCREB_PTH30_rep1; Mus musculus; ChIP-Seq
The kidneys were isolated, rinsed with cold 1× phosphate-buffered saline, and subjected to a cross-linking reaction for 10 min with 1.5% formaldehyde. After the fixative was neutralized for 5 min with glycine (0.125mfinal concentration), the kidney tissue was homogenized briefly in phosphate-buffered saline using a loose fitting Dounce pestle, and the cells were collected via 70-μm nylon cell strainers (BD Falcon). Cells were then subjected to ChIP analysis. ChIP-seq libraries were prepared using the NEBNext Ultra II DNA kit (NEB, #E7645S) with the NEBNext Multiplex Oligos for Illumina (NEB, #E6440S) according to the manufacturer's protocols. Libraries were submitted to Azenta/Genewiz and sequenced on a NovaSeq X. Paired-end, 150 bp sequencing with a target of 30+ million reads was performed.