GSM4711132: Primary islets CTL-1; Homo sapiens; RNA-Seq
Total RNA extraction were performed with Rneasy-micro plus kit based on Qiagen's instruction. Libraries were prepared according to instructions accompanying Takara's SMART-Seq v4 for ultra-low RNA inputs kit (R400752). Briefly, Upto 10 ng of sample was prepared using Takara's SMART-Seq v4 Ultra Low Input RNA protocol per manufacturer's guidelines. Briefly, RNA underwent reverse transcription via template switching and amplification, resulting in double stranded cDNA. The amount of cDNA between 100-3000 bp was calculated by Fragment Analyzer and Qubit. 150pg of the sample in range was then added to Illumina's Nextera XT and processed according to manufacturer's guidelines. Briefly, the cDNA underwent tagmentation, using Nextera XT's transposase, and unique dual indexes were added by PCR amplification. Final libraries went through QC with the Fragment Analyzer and qPCR on a Roche Light Cycler 480 II. All libraries were then pooled and sequenced at single-end 40-50 base-pair using the Illumina HiSeq 2500.
GEO Accession
GSM4711132