GSM6214804: E7.5-DKO-epi-3; Mus musculus; miRNA-Seq
Total RNA was extracted from E6.5 and E7.5 embryos and AG-haESCs by using miRNeasy Micro Kit (QIAGEN, Cat#217084) according to the manufacturer's instructions. The preparation of CAS-seq cDNA library was mainly followed the published protocol (Yang. Q., et al., 2019, Nature Communications). The extracted RNA (100 pg) underwent the following steps to construct small RNA libraries: adapter ligation, first-strand cDNA synthesis, Cas9/sgRNA treatment, on-bead in vitro transcription, second RT and PCR amplification and gel purification. An Illumina NovaSeq 6000 System was used to sequence the small RNA libraries at the MingmaTechnologies Co., Ltd., China