title: GSM2869016: scRNAseq_072_cell_13; Mus musculus; RNA-Seq
library_construction_protocol: For single cell RNA sequencing, 4000 cells were loaded onto a 10-17 μm Fluidigm C1 Single Cell mRNA Seq IFC, and cells captured according to manufacturer's instructions. The captured cells were manually annotated under a microscope, to identify capture chambers containing cell debris, dead cells, no cells or multiple cells. Reverse transcription and cDNA preamplification was then performed in the 10-17 μm Fluidigm C1 Single Cell mRNA Seq IFC using the SMARTer Ultra Low RNA Kit for the Fluidigm C1 System and Advantage 2 PCR kit. cDNA was harvested and diluted to 0.1-0.3 ng/μl. Sequencing libraries were then prepared using the Nextera XT DNA Sample Preparation Kit and Nextera XT DNA Sample Preparation Index Kit according to the Fluidigm instructions, “Using the C1™ Single-Cell Auto Prep System to Generate mRNA from Single Cells and Libraries for Sequencing.” The 96 libraries from one IFC were pooled and 150 bp paired-end sequencing performed on the Illumina HiSeq2000.
attributes.tag: GEO Accession
attributes.value: GSM2869016