GSM8648703: Pol3 CUT&Tag, mESCs, KO, preCond, HS0, rep2; Mus musculus; OTHER
CUT&Tag was performed on 100,000 cells with 1:10 diluted primary antibody, 1:100 diluted secondary antibody, and 1:20 diluted in house produced pA-Tn5. DNA was released from the cells with an overnight proteinase K digestion at 37C, followed by phenol-chloroform extraction. Libraries were constructed using 21 µL of DNA, 2 µL Universal i5 primer (10 µM), 2 µL uniquely barcoded i7 primer, and 25 µL NEBNext HiFi 2x PCR Master mix. 12 cycles of PCR was used to amplify the CUT&Tag libraries, followed by DNA cleanup using SPRI beads.