title: GSM8994439: 100 embryos_4SU_rep1; Mus musculus; OTHER
library_name: GSM8994439
library_construction_protocol: For SLAM-seq, the collected embryos were washed six times with 1xPBS and then immediately placed in lysis buffer (Combine 0.2% (vol/vol) Triton X-100 and 2 U µl–1 RNase inhibitor in a volume of 1 µl) and then frozen at -80℃;  For ATAC-seq, the collected embryos were lysed in 5 µl lysis buffer (10 mM Tris-HCl (pH 7.5), 10 mM NaCl, 3 mM MgCl2, and 0.01% digitonin) for 10 min on ice. For SLAM-seq, samples were taken out from -80℃ freezer, and thawed on ice, 1 µl of alkylation mix was added to the lysis buffer for a final alkylation reaction of 50 mM Tris-HCl (pH 8.4), 10 mM IAA, 45% DMSO. After 15 min at 50 °C, the alkylation reaction was stopped by adding 2 µl of stop solution for a final alkylation reaction of 15 mM DTT, 2 mM dNTP mix (Thermo Fisher Scientific, R0192), 10 µM oligo-dT primer and 2 U µl–1 RNase inhibitor, incubating for 5 min at room temperature. Reverse transcription (RT) and the remaining library preparation were performed according to a modified version of Smart-seq2. The modification step was the removal of DTT from the RT mix. Full length cDNA libraries were fragmented and processed into sequencing libraries by using the TruePrep DNA Library Prep Kit for Illumina (Vazyme, TD503) following the manufacturer's instructions; For ATAC-seq, tagmentation and remaining library preparation were performed by using the Hyperactive ATAC-Seq Library Prep Kit for Illumina (Vazyme, TD711) with minor modification. The tagmentation mix were modified by adding 2.5 µl TTE Mix V50.