title: GSM3833256: KO-4cell-3; Mus musculus; RNA-Seq
library_construction_protocol: 2-5embryos were collected as one replicate. Zona pellucida of embryos were removed with 0.5% protease E (Sigma). Embryos were washed for 5-6 times in 0.5% BSA(Sigma) in PBS solution before put into cell lysis buffer. After incubation at 70°C for 90 seconds, reverse transcription for the first strand cDNA was performed subsequently with a 24 oligo (dT) primer. After second strand cDNA synthesis, cDNA library was amplified for 16-18 cycles based on the embryo number. Amplified cDNA was diluted for 10 times for qPCR reaction. For library construction, amplified cDNA was sheered to 200-300 bp using Covaris S220 system. The sequence libraries were generated using KAPA HyperPlus Library Preparation kit (Roche, KK8510), following the manufacturer's instructions. 8-12 cycles amplification was used depending on cDNA concentration.
attributes.tag: GEO Accession
attributes.value: GSM3833256