GSM2756391: T3_no_PFA_Control_a; Mus musculus; RNA-Seq
Total RNA was extracted from fixed MuSCs using the RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE, Ambion, AM1975. Modification: cells were incubated at 50°C for 1h in digestion buffer plus Proteases. No 80°C incubation was performed as recommended by the manufacturer as this deteriorated RNA quality. Directional libraries were prepared using the Smarter Stranded Total RNA-Seq kit- Pico Input Mammalian following manufacturer's instructions (Clontech, 635005). The quality of all libraries was assessed with the DNA-1000 kit (Agilent) on a 2100 Bioanalyzer and quantification was performed with Quant-It assays on a Qubit 3.0 fluorometer (Invitrogen). Clusters were generated for the resulting libraries, with Illumina HiSeq SR Cluster Kit v4 reagents. Sequencing was performed with the Illumina HiSeq 2500 system and HiSeq SBS kit v4 reagents. Runs were carried out over 65 cycles, including seven indexing cycles, to obtain 65-bp single-end reads. Sequencing data were processed with Illumina Pipeline software (Casava version 1.9).
GEO Accession
GSM2756391