title: GSM7754907: D0_DnmtTKO_H3K36me3_rep2; Mus musculus; OTHER
library_construction_protocol: We performed CUT&Tag according with the protocol described in (Kaya-Okur et al., 2020), closely following the procedure described in “Bench top CUT&Tag V.2” (S Kaya-Okur and Henikoff, 2019). 500,000 cells were used for each sample and duplicates were performed. The primaries antibodies, anti-H3K27me3 (Cell signal C36B11) 1:100, the anti-H3K36me3 (Invitrogene RM491) 1:100, were incubated overnight at 4ºC on a rotator. Samples were then incubated with a secondary anti-rabbit IgG (ABIN6923140) at room temperature on a rotator.  After an incubation of 1h at room temperature with a pA-Tn5 adapter complex (Addgene #124601), the Tn5 enzyme tagmentation reaction was performed by adding a buffer containing MgCl2 to the samples and incubating for 1h at 37°C. To stop tagmentation and solubilize DNA fragments, we added 10 μL 0.5M EDTA, 3 μL 10% SDS and 2.5 μL 20 mg/mL Proteinase K to each sample and samples were incubated at 50°C for 1h. Purified DNA was obtained by phenol/chloroform extraction and precipitated with 100% ethanol and centrifugation. The DNA pellet was washed in 80% ethanol, spun down and air-dried before being resuspended in 25µl of 1mM Tris-HCl pH8.0. Nextera primers or indexed primers were used to perform PCR. Library quality was assessed using 4200 TapeStation system (Agilent) and paired-end, 150 bp sequencing on an Illumina Novaseq was performed by Novogene Co.