GSM1974093: 11a-wt1-WT; Mus musculus; RNA-Seq
For RNA extraction brain tissue was lysed in Qiazol solution (Qiagen) using TissueLyser II (Qiagen) tissue homogenizer  follow by RNA isolation using the NucleoSpin RNA Midi kit (Macherey-Nagel). The Agilent TapeStation was used to assess RNA quality. Samples with RINs (RNA Integrity Numbers) of eight or greater were prepped using the Illumina TruSeq mRNA Sample Prep v2 kit (Catalog #RS-122-2001 and RS-122-2002). mRNA was isolated from 0.1-3 ug of total RNA by polyA+ purification, fragmented, and copied into first-strand cDNA using reverse transcriptase and random primers. 3' cDNA ends were then adenylated and adapters ligated. One of the adapters contained a 6-nucleotide barcode to enable multiplexing of samples. Products were purified and enriched by PCR to create the final cDNA library. Libraries were checked for quality and quantity by Agilent TapeStation and qPCR using a library quantification kit for Illumina sequencing platforms (catalog #KK4835, Kapa Biosystems, Wilmington, MA). Clonal clusters were generated using cBot (Illumina, Inc., San Diego, CA). Quadriplexed samples were sequenced using the HiSeq 2000 system (Illumina) with a 100-cycle paired-end run in high output mode using Version 3 reagents according to manufacturer's protocols.