C4
Homo sapiens
Protocols: Forty milliliters of peripheral blood was collected from a healthy-steady state donor. Peripheral blood mononuclear cells (PBMCs) were isolated with a lymphoprep gradient following B cell enrichment with rosetteSep (stem cells 15064). Cells were stained for isolation via fluorescence activated cell sorting; CD19 (BioLegend, clone H1B19), CD27 (BioLegend, clone O323), CD38 (BioLegend, clone HIT2), IgG (BD, clone G18-145), IgA (Southern Biotech, clone G1012-YG02B) and IgM (Southern Biotech, clone UHB). Plasmablasts, CD19+CD27++CD38++IgA+, were bulk and then single cell sorted on a BD FACS AriaII. A successful purity check was performed at the time of plate sorting. 'Libraries were prepared according to the published SmartSeq2 protocol. Briefly, cells were sorted directly into lysis buffer containing an oligo designed to anneal to the poly-A tails of mRNA. Reverse transcription was performed with Invitrogen''s Superscript III enzyme (cat#18080-044). Three different pre-amplification PCR cycle numbers were tested and 3ul of the amplified cDNA was removed for the PCR based antibody amplification process. The Nextera XT DNA sample perparation kit (cat#FC-131-1024) was used for the tagmentation and indexing reactions, performed with .5ng of the amplified cDNA.'
ENA first public
2016-06-02
ENA last update
2016-05-23
External Id
SAMEA4002745
INSDC center alias
Toyota Technological Institute at Chicago
INSDC center name
Toyota Technological Institute at Chicago
INSDC first public
2016-06-02T17:01:41Z
INSDC last update
2016-05-23T14:35:35Z
INSDC status
public
Submitter Id
E-MTAB-4745:C4
age
27
broker name
ArrayExpress
cell type
IgA plasmablast
common name
human
organism part
blood
sample name
E-MTAB-4745:C4
sex
female