GSM8547209: 2C.FLAG.2; Mus musculus; RIP-Seq
400 2-cells for each sample were washed three times with 0.2% bovine serum albumin (BSA) in PBS and lysed in the lysis buffer (50 mM Tris–HCl [pH 7.4], 1% Triton X‐100, 150 mM NaCl, 5 mM EDTA, protease inhibitor cocktail, and RNase inhibitor). 90% cell lysate supernatant was used for immunoprecipitation with agarose beads conjugated with IgG or FLAG antibodies, while the residual cell lysate supernatant was used as the “input”. After incubation at 4°C for 4 h, bead‐bound RNAs were collected and reverse‐transcribed using the random primer. The samples were reverse transcribed and amplificated using the Smart-seq2 method, as described previously. Sequencing libraries were constructed from 500 pg of amplified cDNA using TruePrep DNA Library Prep Kit V2 for Illumina (Vazyme, TD503). Barcoded libraries were pooled and sequenced on the HiSeq X Ten platform with 150-bp paired-end reads.