GSM3262427: mouse1; Mus musculus; RNA-Seq
RNA isolation, (q)RT-PCR and RNA-Seq - Total RNA was isolated using Tri reagent (Sigma)  and digested by DNase I (Life Technology). First-Strand cDNA was synthesized using  SuperScript III reverse transcriptase (Life Technology) according to manufacture's protocol and  standard PCR was performed afterwards. Quantitative Real-time PCR analysis was  accomplished using Fast SYBR Green master mix on a 7500 quantitative Real-time PCR  machine (Applied Biosystems). The relative amount of gene transcripts and pre-micro-RNA  (pre-miRNA) was normalized to GAPDH and U6, respectively. 350 to 900 ng of total RNA determined by Invitrogen Qbit high sensitivity spectrofluorometric measurement is poly A selected and reverse transcribed using Illumina's TruSeq RNA library preparation kit V2. Each sample can be fitted with one of 24 adapters containing a different 6 base molecular barcode for high level multiplexing. After 12 cycles of PCR amplification, completed libraries are sequenced on an Illumina HiSeq2000 or HiSeq2500, generating around 20-50 million high quality 50 base long reads per sample.
GEO Accession
GSM3262427