title: GSM3034728: WT 1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted with RNeasy Mini kit according to manufacturer's protocol (Qiagen, Germantown, MD). RNA libraries were prepared according to the manufacturer's instructions for the TruSeq RNA Sample Prep Kit v2 (Illumina, San Diego, CA) from 100 ng of total RNA. Briefly, polyA mRNA was purified from total RNA using oligo dT magnetic beads. Purified mRNA was fragmented at 95°C for 8 min and eluted from the beads. Double stranded cDNA was prepared using SuperScript III reverse transcriptase, random primers (Invitrogen, Thermo Fisher Scientific, Waltham, MA) and DNA polymerase I and RNase H. The cDNA ends were repaired and an “A” base added to the 3' ends. TruSeq paired end index DNA adaptors (Illumina, San Diego, CA) with a single “T” base overhang at the 3' end were ligated and resulting constructs were purified using AMPure SPRI beads (Agencourt Bioscience, Beverly, MA). The adapter-modified DNA fragments were enriched by 12 cycles of PCR using Illumina TruSeq PCR primers (Illumina, San Diego, CA). The concentration and size distribution of the libraries were determined using an Agilent Bioanalyzer DNA 1000 chip (Agilent Technologies, Santa Clara, CA) and Qubit fluorometry (Invitrogen, Thermo Fisher Scientific, Waltham, MA).
attributes.tag: GEO Accession
attributes.value: GSM3034728