GSM6503259: 33hpi_control_rep1; Mus musculus; RNA-Seq
For RNA-seq, the collected embryos were washed twice with 1xPBS and then immediately placed in lysis buffer( Combine 0.2% (vol/vol) Triton X-100 and 2 U µl–1 RNase inhibitor);For Stacc-seq, embryos were   collected and transferred into a 200-μl low-binding tube (total volume with buffer less than 1 μl). Six microlitres of DB-1  (10 mM Tris-HCl pH = 7.4, 150 mM NaCl, 0.5 mM spermidine, 1 × EDTA-free Roche complete protease inhibitor, 0.005% digitonin (Sigma, D141)) buffer was added. For RNA-seq, samples were taken out from -80℃ freezer, and thawed on ice, 1μl of 10μM oligo-dT primer, 1μl of 10 mM dNTP mix were added to the lysis buffer. cDNA synthesis and amplification were performed following the Smart-seq2 ; Stacc-seq was performed following the previously reported protocol designed by Bofeng Liu etc. pG–Tn5 (Vazyme,TD901) and RNA pol ll antibody (Active motif,39097,lot102660) was used. The precipitated DNA was processed into sequencing libraries using the Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme,TD903 )following the manufacturer's instructions.