GSM2200201: P1 Cochlea GFP- 2; Mus musculus; RNA-Seq
Total RNA was extracted using Trizol (Thermo Fisher Scientific) and PelletPaint (Novagen EMD Millipore). Briefly, samples were homogenized in 300µl of Trizol and shaken with 60µl chloroform and 2µl PelletPaint and centrifuged for 15 minutes. The supernatant containing RNA was precipitated with 1 volume of isopropanol, centrifuged for 30 minutes, washed in 75% ethanol, resuspended in 100µl water and re-precipitated with 10µl 3M sodium acetate and 250µl ethanol. After a final wash in 75% ethanol, the pellet was allowed to dry, resuspended in 20µl RNAse-free water and stored at -80°C. cDNA libraries for RNAseq were generated using RNA Seq Truseq RNA sample preparation kit v2 (Illumina) following the “low sample” protocol according to the manufacturer's instructions for mRNA extraction, cDNA synthesis, indexing and amplification. cDNA generated from less than 20,000 cells received an initial amplification using the NuGen Ovation Kit.  The quality and integrity of RNA samples and the final quality of the sequencing libraries was checked by electrophenogram in an Agilent Bioanalyzer. paired end 100bp sequencing
GEO
GSM2200201