GSM1303926: islet preparation 1 under control condition; Homo sapiens; RNA-Seq
Total RNA was isolated using the RNeasy Mini Kit (Qiagen, Venlo, The Netherlands), which favors purification of all RNA molecules longer than 200 nucleotides and sample preparation was done as described by the manufacturer (Illumina, Eindhoven, The Netherlands). Briefly, mRNA was purified from two μg total RNA using oligo (dT) beads, before it was fragmented and randomly primed for reverse transcription followed by second-strand synthesis to create ds cDNA fragments. The generated cDNA had undergone paired-end repair to convert overhangs into blunt ends. After 3’-monoadenylation and adaptor ligation, cDNAs were purified on a 2% agarose gel and 200 basepair (bp) products were excised from the gel. Following gel digestion, purified cDNA was amplified by PCR using primers specific for the ligated adaptors. The generated libraries were submitted to quality control with the Agilent bioanalyzer 2100 (Agilent Technologies, Wokingham, UK) before sequencing. The RNA integrity number (RIN) values for all samples were 7.5 and above. 1 μL cDNA was loaded on an Agilent DNA chip (DNA-1000) to verify cDNA quality and quantity. Only libraries reaching satisfactory conditions were used for sequencing, on one sequencing lane of an Illumina Genome Analyzer II system (GAII, Illumina).
GEO Accession
GSM1303926