GSM2752505: VMP_Tag_seq; Rattus norvegicus; RNA-Seq
Urogenital tracts were removed from day of birth Wistar rats and microdissected into VMP and SU components using a Leica MZ6 dissection microscope. For RNA-sequencing only, VMP and SU tissues were disscociated with 2mg/mL Collagenase 1A (Sigma-Aldrich, Missouri, USA) for 60 minutes at 37 degrees C. Dispersed fibroblasts were separated by centrifugation and passing through a 0.7um cell strainer (Falcon, Corning, New York, USA) Microdissected tissues (VMP and SU) were processed for digital gene expression Tag-profiling using NlaIII and a protocol provided by Illumina followed by sequencing on an llumina GAIIX (1x 50 SE) at one lane per sample (25-30m reads). PolyA+ RNA was purified, cDNA synthesised, digested with NlaIII and ligated to Adaptor 1 (containing an Mme1 site). Samples were digested with Mme1, ligated to Adaptor 2, and PCR amplified, followed by gel electrophoresis and purification of 85bp fragments that were sequenced
GEO Accession
GSM2752505