Cut&Tag-Kikue-4H8-NSN-4H8-rep1 (CRX2069181)
50 to 80 mouse oocytes (per sample) were pre-extracted with ice-cold extraction buffer [25 mM HEPES (pH 7.4), 50 mM NaCl, 3 mM MgCl2, 300 mM sucrose and 0.5% triton X-100] for 10 min on ice and washed three times through ice-cold extraction buffer without triton X-100. Pre-extracted oocytes were directly washed with antibody buffer and incubated with primary antibody without crosslinking. Primary antibodies used were mouse IgG (Beyotime #A7028), mouse anti-CTD pS5 (4H8) (Santa Cruz #sc-47701), rat IgG (Beyotime #A7031), rat anti-CTD pS2 (Active motif #61699). Secondary antibodies used were rabbit anti-mouse (Sigma-Aldrich #06-371) and rabbit anti-rat (Sigma-Aldrich #AP164). In the library amplification step, 16 to 21 cycles were used. The purified libraries were quantified by Qubit 1X dsDNA HS Assay Kit and qPCR, analyzed by a Fragment Analyzer System to confirm successful amplification and quality, and sequenced using Novaseq 6000 Sequencing System.