title: GSM3384439: RNA-seq.siZmym2_normal_8cell_0; Mus musculus; RNA-Seq
library_construction_protocol: For ESCs, total RNAs were extracted using the RNeasy kit (#74136, Qiagen) according to the manufacturer's instructions. For embryos, a total of 10-20 embryos with the same treatment and embryonic stage were pooled for each reaction. Injected embryos were harvested, washed several times in 0.5% BSA-PBS (Sigma) solution and subsequently picked and transferred into lysis buffer by a mouth pipette. For ESCs, 1 µg total RNA from each sample was taken for the preparation of PolyA RNA-seq libraries, and massively parallel sequencing was performed with the HiSeq4000 platform. Paired-end 150 bp-length reads were produced. For embryos, RNA-seq libraries were prepared according to the Smart-seq2 protocol (Picelli et al., 2014) with minor modifications. After reverse transcription, cDNA was pre-amplified (10 PCR cycles) and purified with Ampure XP Beads (Agencourt) at 0.8 beads/1 DNA (v/v). One microliter (1 μl) cDNA, dulited by 19 μl nuclease-free water, was used for Real-time PCR quality check. The amplified cDNA was fragmented using a Covaris sonicator (Covaris S220). To generate the sequence libraries, the KAPA Hyper Prep Kit was used following the manufacturer's instructions. NOVA pair-end 150-bp sequencing was performed on a HiSeq 2500 sequencer (Illumina) at Berry Genomics Corporation.
identifiers.namespace: GEO
identifiers.id: GSM3384439