GSM2482651: 32 FF; Mus musculus; RNA-Seq
Trizol extraction of total RNA was performed according to the manufacturer's protocol followed by purification on a silica-gel-based membrane (Rneasy-Mini; Qiagen, Valecia, CA). Libraries were constructed from first strand cDNA using ScriptSeqTM v2 RNA-Seq library preparation kit. Briefly, 50ng of rRNA-depleted RNA was fragmented and reverse transcribed using random primers containing a 5' tagging sequence, followed by 3'end tagging with a terminal-tagging oligo to yield di-tagged, single-stranded cDNA. Following purification by a magnetic bead-based approach, the di-tagged cDNA was amplified by limit-cycle PCR using primer pairs that anneal to tagging sequences and add adaptor sequences required for sequencing cluster generation. Amplified RNA-seq libraries were purified using AMPure XP System.
GEO Accession
GSM2482651