Sample 62
Protocols: Peritoneal lavage was collected following the instructions described previously43. Briefly, ice-cold DPBS was injected into the abdominal cavity. After massaging the peritoneum, the peritoneal fluid was collected by a needle with a syringe. The lavage was centrifuged at 400 x g for 7 min. Lysing Buffer (Thermo Scientific). Peritoneal B cells were enriched with the CD19 MicroBeads (Miltenyi Biotech). C57BL/6 wild-type mice were used for scRNA-seq and scBCR-seq. The Bcl11aeGFP mice were previously described42. The Rag2-/- mice (008449) were purchased from the Jackson Laboratory. All mice used were maintained under a specific pathogen-free condition at the Laboratory Animal Research Center of Tongji University. All experiments were approved by the Biological Research Ethics Committee of Tongji University. scRNA-seq libraries were prepared following the SMART-seq2 protocol44. Single cells were directly sorted into 96-well plates containing the lysis buffer and external RNA spike-ins (Ambion) (1:500,000). The reverse transcription and pre-amplification were performed, then the amplified cDNA products were purified. Quality control was performed by checking the size distribution of cDNAs. Multiplex (96-plex) libraries were constructed and amplified using the Nextera XT library preparation kit (Illumina). The libraries were then purified and the quality of the library is assessed by the Bioanalyzer (Agilent) before submission to the DNA sequencing pipeline.
ENA first public
2021-08-14
ENA last update
2021-08-14
External Id
SAMEA8061126
INSDC center alias
EMBL-EBI
INSDC center name
EMBL-EBI
INSDC first public
2021-08-14T00:12:13Z
INSDC last update
2021-08-14T00:12:13Z
INSDC status
public
Submitter Id
E-MTAB-10061:Sample 62
age
8.1
broker name
ArrayExpress
cell type
B-1 B cell
common name
house mouse
developmental stage
adult
genotype
wild type genotype
individual
SCGC--2254
organism part
peritoneal fluid
sample name
E-MTAB-10061:Sample 62
sex
female
single cell identifier
SCGC--2254_F02
strain
C57BL/6