GSM5848733: Lats1-CKO luminal, BM7, ATAC-seq; Mus musculus; ATAC-seq
Nuclei were incubated with 2ul of Nextera Tn5 enzyme (TDE1, Illumina) for 1h at 37°C. Enzyme inactivation was done by addition of 5ul Clean-up buffer (900mM NaCl, 30mM EDTA), 2ul of 5% SDS and 2ul of Proteinase K (NEB) and incubation for 30min at 40°C. Tagmented DNA was isolated using 2x SPRI beads cleanup. two sequential 9-cycle and 5-cycle PCR were performed in order to enrich small tagmented DNA fragments. Libraries were prepared using KAPA HiFi HotStart ready mix. After the first PCR, the libraries were selected for small fragments using SPRI cleanup (0.65x). Then a second PCR was performed with the same conditions in order to obtain the final library.