GSM3036766: WT PAX6-TBR2- cells, replicate 2; Mus musculus; RNA-Seq
Sorted cells were resuspended in the lysis solution of RecoverAll total nucleic acid isolation kit (Ambion) and incubated for 3 hrs at 50oC. RNA was then purified according to manufacturer's instructions. The concentration and quality of purified RNA was determined using BioAnalyzer (Agilent) RNA was reverse-transcribed into cDNA and amplified by RNA-based single primer isothermal amplification (SPIA) using the Ovation RNA-seq system V2 (NuGEN). Synthesized cDNA was sonicated using a Covaris S2 ultrasonicator to reduce fragment size range to 100-600 bp. For library preparation, 100 ng of sheared cDNA was end repaired, ligated with barcoded adaptors, amplified for 9 PCR cycles and purified using the Ovation Ultralow System V2 (NuGEN).
GEO Accession
GSM3036766