title: GSM4162266: cKO 12h 0Gy I18_1045_02_3865; Mus musculus; RNA-Seq
library_construction_protocol: Fetal cortices were dissected and stored in RLT Plus lysis buffer (Qiagen) at -80°C until RNA extraction using the RNeasy Mini Kit (Qiagen). RNA quality was determined using the RNA nano assay on a 2100 Bioanalyzer (Agilent Technologies). All samples had RNA Integrity Numbers >9.10. Triplicate RNA-Seq libraries were prepared according to the TruSeq stranded mRNA protocol (Illumina). Briefly, 200 ng of total RNA was purified using poly-T oligo-attached magnetic beads to end up with poly-A containing mRNA. The poly-A tailed mRNA was fragmented and cDNA was synthesized using SuperScript II and random primers in the presence of Actinomycin D. cDNA fragments were end repaired, purified with AMPure XP beads, A-tailed using Klenow exo-enzyme in the presence of dATP. Paired end adapters with dual index (Illumina) were ligated to the A-tailed cDNA fragments and purified using AMPure XP beads. The resulting adapter-modified cDNA fragments were enriched by PCR using Phusion polymerase as followed: 30 s at 98°C; 15 cycles of 10 s at 98°C, 30 s at 60°C, 30 s at 72°C; 5 min at 72°C. PCR products were purified using AMPure XP beads and eluted in 30 µl of resuspension buffer. One microliter was loaded on an Agilent Technologies 2100 Bioanalyzer using a DNA 1000 assay to determine the library concentration and for quality check.
attributes.tag: GEO Accession
attributes.value: GSM4162266