SP_replicate1_cell_119
Mus musculus
Protocols: Pdgfra+ Sca1+ Cd31- cells were isolated using FACS sorting from ventricles of male adult mice. Briefly, hearts were minced and incubated in collagenase type II (Worthington, USA) at 37°C before to be filtered on 40μm strainers. Cells were then resuspended in red cell lysis buffer followed by dead cell removal and immunostaining for 15min on ice with fluorophore-conjugated antibodies and followed by FACS sorting for mentioned markers. Immediately after cell sorting and counting, the cells were loaded on the integrated fluidics circuits (IFC) C1 chip. A LIVE/DEAD cell staining kit (Life Technologies, USA) was used to mark live and dead cells with green (live) and red (dead) fluorescence before the run. Each capture site was carefully examined under a Leica fluorescence microscope in bright field, Red, and Green fluorescence channels for cell doublets and viability. Finally, 12 µl of cells (200,000 cells/ml) were re-suspended in 8 microliters of suspension reagent (Fluidigm), of which 7 microliters could be loaded on the Fluidigm Single Cell Auto Prep IFC optimized for 5-10 μm cell-size. The 96 trapping sites on the chip were visualized by the microscope to ensure the capture rate was satisfactory, before cell lysis and cDNA preparation. The reverse transcription was performed using Clontech SMARTer Ultra Low Input RNA Kit V4 (Takara-Clontech, USA) on the chip. SMARTer V3 Nextera XT protocol from Illumina
ENA-FIRST-PUBLIC
2019-03-28T17:03:51Z
ENA-LAST-UPDATE
2018-11-06T14:35:53Z
External Id
SAMEA5066718
INSDC center name
Victor Chang Cardiac Research Institute
INSDC first public
2019-03-28T17:03:51Z
INSDC last update
2018-11-06T14:35:53Z
INSDC status
public
Submitter Id
E-MTAB-7365:SP_replicate1_cell_119
age
8
broker name
ArrayExpress
common name
house mouse
developmental stage
adult
disease
normal
genotype
wild type genotype
organism part
cardiac ventricle
phenotype
Pdgfra+/Sca1+/Cd31-
post analysis well quality
pass
sample name
E-MTAB-7365:SP_replicate1_cell_119
scientific_name
Mus musculus
sex
male
single cell quality
OK
single cell well quality
OK
strain
C57BL/6J