GSM5089935: IP_WT_II; Mus musculus; OTHER
~20 mio cells were harvested from a 15 cm dish and genomic DNA was extracted with the genomic tips kit 100 (Qiagen). 50 ug of DNA were subjected to SSB-seq protocol. Briefly: Nick-translation was performed with E-coli Pol I at 16°C for 1 min, including DIG-dUTPs and ddNTPs to prevent exceeding polymerization. The labelled DNA was precipitated, purified and subsequently sonicated and digested with restriction enzymes to get rid of high-molecular DNA. The fragmented DNA was purified again and subjected to immunoprecipitation using an anti-DIG antibody (Roche 11333089001) and Protein G Dynabeads (Life Technologies). Immunoprecipitated DNA was purified with the ChIP DNA clean & concentrator kit (Zymo). 10 ng of each immunoprecipitated DNA and Input DNA were subjected to library construction with a KAPA HyperPrep Kit (Roche) using 8 cycles of amplification.