GSM5906373: Input rep 1; Mus musculus; ChIP-Seq
E16.5 kidney chromatin was extracted from isolated nuclei and sheared to 100-1000bp. Primary antibody was incubated overnight for ChIP reactions. Crosslinks were reversed by adding NaCl and heating at 65 °C for 15 h, RNase A was added and incubated for 2 h at 37 °C, and Proteinase K added and incubated 2 h at 63 °C. DNA was isolated using the MinElute PCR Purification Kit (Qiagen 28004). 2 ng DNA was sheared to 100–300 bp. Libraries were constructed using the Ion Plus Fragment Library Kit (Thermofisher Scientific 4471269) according to (Dorsett and Misulovin, Methods Mol Biol. 2017; 1515:125-139). 