alias: GSM1677296
title: GSM1677296: DMSO_66; Rattus norvegicus; RNA-Seq
library_construction_protocol: Following quantification, dispersed islets were loaded into Fluidigm C1 IFC microfluidic chips according to manufacturer specifications (Fluidigm). Briefly, 60 µL of a 200 cell/µL mixture was combined with 40 µL of C1 suspension reagent. 5 µL was loaded into the 10-17 µm Fluidigm IFC using the mRNA Seq: Cell Load (1771x/1772x/1773x) protocol. After loading, the IFC was manually inspected to ensure single cell capture by phase contrast microscopy. Following inspection, cells were lysed according to Fluidigm's C1 protocol. Reverse transcription, and PCR amplification were all performed on the IFC using the SMARTer Ultra Low RNA kit (Clontech) with the mRNA Seq: RT & Amp (1771x/1772x/1773x) protocol. cDNA was quantified by Pico Green assay (Life Technologies) and then normalized with C1 DNA dilution buffer to 0.1-0.3ng/µL.  Sequencing libraries were prepared with the Nextera XT DNA Sample Preparation Index Kit, (Illumina, PN FC-131-1002).  Sequencing libraries were run on an Illumina HiSeq 1000 sequencer using single 50 bp reads, one 96-well plate per lane.
identifiers.namespace: GEO
identifiers.id: GSM1677296