Mus musculus
Protocols: Tissue of male C57BL6J mice was harvested, digested, and stromal cells isolated via differential centrifugation. The lineage-negative fraction of stromal cells was purified via affinity column. Following centrifugation and removal of the medium, cells were resuspended at a concentration of 150–500 cells/μL. This cell suspension was mixed with C1 Cell Suspension Reagent (Fluidigm, Cat # 634833) at the recommended ratio of 3:2 immediately before loading 5 μL of this final mix on the C1 IFC. Cells were lysed via Module 2 mRNA-seq kit per Fluidigm protocol specifications (PN 100-7168). Resultant mRNA was stabilized and reverse-transcribed using the SMARTer kit (Clontech), also per Fluidigm protocol specifications. Approximate run time was ~7.75 hours (6.5 hours for lysis, reverse transcription, and amplification; and 1.25 hours for harvest). 3µL amplicons were diluted with 10µL C1 DNA Dilution Reagent (Fluidigm) to generate 13 µL mix, and transferred to a new plate. Sample concentrations were normalized manually, then prepared for successive tagmentation, amplification, and pooling per the Illumina Nextera XT DNA Library Preparation protocol guidelines.
ENA-FIRST-PUBLIC
2019-12-23T04:03:07Z
ENA-LAST-UPDATE
2019-11-14T10:12:29Z
External Id
SAMEA6225039
INSDC center name
UCSF
INSDC first public
2019-12-23T04:03:07Z
INSDC last update
2019-11-14T10:12:29Z
INSDC status
public
Submitter Id
E-MTAB-8464:Epi_(no-Tx)_single-23
age
10
broker name
ArrayExpress
cell type
stromal cell
common name
house mouse
developmental stage
adult
genotype
wild type genotype
immunophenotype
Lin-
organism part
epididymal adipose tissue
sample name
E-MTAB-8464:Epi_(no-Tx)_single-23
scientific_name
Mus musculus
sex
male
strain
C57BL/6J