title: GSM5994255: IVF_2c_RNAseq; Capra hircus; RNA-Seq
library_construction_protocol: Total RNA was isolated from in vitro fertilization (IVF) goat embryos at the two-cell and eight-cell stages and somatic cell nuclear transfer(SCNT) 8-cell embyos. We used the Qubit® RNA Assay Kit and Qubit® 2.0 Fluorometer (Life Technologies, CA, USA) to detect RNA concentration. RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies, CA, USA) was employed for RNA integrity evaluation, and SMART-SeqTM v4 UltraTM Low RNA Kit for Sequencing (Clontech) was used to obtain the amplified cDNA in accordance with the manufacturer's instructions. The Covaris system sheared the cDNA samples prior to library preparation. Moreover, sequencing libraries (IVF-2c  IVF-8c and SCNT-8c) were generated using NEBNext® UltraTM DNA Library Prep Kit for Illumina® (NEB, USA) in accordance with the manufacturer's instructions, and index codes were added to attribute sequences to each sample. Clustering of the index-coded samples was conducted on a cBot Closter Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumina) in accordance with the manufacturer's instructions. After cluster generation, library preparations were sequenced on an Illumina HiseqTM 2500 platform, and 125-bp paired-end reads were generated.