title: GSM3383076: 20140414_C81_150331_3; Mus musculus; RNA-Seq
library_construction_protocol: XX gonads were collected in PBS at room temperature. PBS was removed and tissues were digested 10 minutes at 37°C with trypsin/EDTA 0.05% and filtered through a 40µm cell strainer to obtain single cell suspension. Nr5a1-eGFP+ cells were then sorted by fluorescent-active cell sorting (BD FACSARIA II) with Draq7TM dye to exclude dead cells and collected in PBS for the single-cell capture. Cells were centrifuged at low speed to be concentrated at an average of 400 million cells per mL prior cell capture. Single cells were capture on small size IFC microfluidic chips using the C1 Single-Cell Auto Prep System (Fluidigm) following the manufacturer recommended protocol. Reverse-transcription and pre-amplification of the single-cell cDNAs were achieved within the IFC chip using the SMARTer Ultra Low RNA kit for Illumina (Clontech) according to the C1 protocol. RNA-sequencing libraries of the single-cell cDNA were prepared using the Illumina Nextera XT DNA Sample Preparation kit using the modified protocol described in the C1 documentation.
identifiers.namespace: GEO
identifiers.id: GSM3383076