GSM3618016: PUB33; Mus musculus; RNA-Seq
Total pancreatic RNA was isolated using guanidine thiocyanate, followed by acid phenol-chloroform extraction. 1µg of total RNA samples, each containing an equal amount of ERCC ExFold RNA Spike-In Mix 1 (Ambion 4456739), was used as provided by the user. Average sample RNA Integrity Number was 8.4 (range 7.8 - 9.2) when assayed on an Agilent 2100 Bioanalyzer. PolyA+ fraction was purified and randomly fragmented, converted to double stranded cDNA and processed through subsequent enzymatic treatments of end-repair, dA-tailing, and ligation to adapters as in Illumina's "TruSeq Stranded mRNA Sample Preparation Part # 15031047 Rev. D" kit (this kit incorporates dUTP during 2nd strand cDNA synthesis, which implies that only the cDNA strand generated during 1st strand synthesis is eventually sequenced). Adapter-ligated library was completed by PCR with Illumina PE primers. The resulting purified cDNA library was applied to an Illumina flow cell for cluster generation and sequenced on an Illumina instrument (see below) by following manufacturer's protocols. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM3618016