GSM6943833: rat Schwann cells - mouse dorsal root ganglia myelinated cocultures treated with DMSO (biol repl 2); Mus musculus; Rattus norvegicus; RNA-Seq
Cell cultures and sciatic nerves were homogenized in TriPure Reagent (Roche) using Precellys Lysing Kit CK14 and homogenizer (8000 rpm, 60 sec at 0°C, twice). RNA extraction was performed according to manufacturer's instruction. RNAseq libraries were prepared from 2500 ng total RNA using the TruSeq RNA Library Preparation Kit v2 (Illumina, San Diego, CA, USA) after poly(A) capture, according to manufacturer's instructions.  Quality and size of RNAseq libraries were assessed by capillary electrophoretic analysis with the Agilent 4200 Tape station (Agilent). Libraries were quantified by real–time PCR against a standard curve with the KAPA Library Quantification Kit (KapaBiosystems, Wilmington, MA, USA). After barcoding, the RNA libraries were pooled, denatured, and diluted to equimolar concentration for further analyses. RNA sequencing was performed by using NextSeq 500 (Illumina) set for 150 cycles in single end (SE). On average 42 million fragments were produced for each sample.