title: GSM3537437: 2%DMSO_rep2; Mus musculus; RNA-Seq
library_construction_protocol: Fifty of 2-cell embryos for each group were collected in 1.5ml tube and then washed with PBS one time. After removing PBS and then the embryos were directly lysed and used for cDNA synthesis. For cDNA synthesis and amplification with low amount of RNA from embryos, we used SMARTer® Ultra® Low Input RNA Kit (Clonetech, 634940) according to the manufacturer's instructions. Using Agilent Bioanalyzer High Sensitivity DNA kit (Agilent, 5067-4626), size and quantity of amplified cDNA were estimated in 400-10,000bp and 2-10ng, respectively. Synthesized cDNA (2ng) was sheared into 150-200 bp size by Covaris™ M220 Focused-ultrasonicator™ (Thermo Fisher Scientific, 4482277) then used for preparation of sequencing library using Low Input DNA Library Prep Kit (Clonetech, 634946) according to the manufacturer's instructions, followed by single end 50 bp sequencing was run on a HiSeq 2500 sequencer (Illumina). Libraries were sequenced on Illumina HiSeq2500 sequencer to generate 50bp PE reads.
identifiers.namespace: GEO
identifiers.id: GSM3537437