GSM3035052
GSM3035052: WTFK1; Mus musculus; RNA-Seq
The RNeasy Micro Kit (Qiagen) was used for the isolation of RNA from organ of Corti cultures. RNA was pooled from 6–8 organ of Corti cultures to obtain a single sample. 200-300 ng of total RNA were reverse transcribed using SuperScript IV reverse transcriptase (Thermo Fisher Scientific), oligo dT primer, and template switching oligo. Second strand cDNA was amplified using LongAmp DNA polymerase (New England BioLabs) for 12 PCR cycles, and purified on SPRI Select Beads (Beckman Coulter). The size distribution and concentration of second strand cDNA was evaluated on a Fragment Analyzer instrument using High Sensitivity Large Fragment Analysis Kit (Advanced Analytical). 125 pg of second strand cDNA was tagmented, PCR amplified, and indexed using the Nextera XT Library Prep Kit (Illumina). Following AMPure XP Bead purification (Beckman Coulter), the size distribution and concentration of cDNA Libraries were evaluated on a Fragment Analyzer instrument using High Sensitivity NGS Fragment Analysis Kit (Advanced Analytical). All libraries were normalized, pooled and sequenced in a HiSeq1500 instrument (126x126 paired end mode).
GEO
GSM3035052
GEO Accession
GSM3035052