GSM2406800: Sox2-deleted undifferentiated NSC_RNA-seq 1; Mus musculus; RNA-Seq
RNA extraction was performed on three independent NSC populations for both wild type and mutant cells, using Trizol and RNeasy Kit (Qiagen). 1/5 volume of chloroform was added to one volume of Trizol. Aqueuos phase was transferred into a new tube. 1.5x volume of ethanol was added and mixed well. Mixture was was filtered through Rneasy (Qiagen) column. Column was washed with Buffer RW1. On-column DNase treatment was performed as described by the RNase-Free DNase Kit (Qiagen). Post treatment column was clean up with Buffer RW1 and two washes of RPE buffer. Column was then dried and total RNA was eluted with RNase-free water. PolyA Stranded Truseq Libraries were generated using the Truseq Stranded mRNA Sample Preparation Kit (Illumina). First, mRNA was purified from 1µg of total RNA using magnetic beads containing poly-T oligos. mRNA was then fragmented and reversed transcribed using Superscript II (Invitrogen), followed by second strand synthesis. Double stranded cDNA was treated with end-pair, A-tailing, adapter ligation and 8 cycles of PCR amplification.
GEO Accession
GSM2406800