Illumina HiSeq 4000 sequencing; The human gastrointestinal tract through space and time- Adult VDJ
The human gastrointestinal tract through space and time- Adult VDJ
For adult samples, fresh mucosal tissue were excised within 60 minutes of circulatory arrest and colon tissue preserved in University of Wisconsin (UW) organ preservation solution (Belzer UW® Cold Storage Solution, Bridge to Life, USA) and mLN stored in saline at 4°C until processing. All tissues were processed immediately after isolation using consistent protocols with variation in enzymatic digestion strength (dx.doi.org/10.17504/protocols.io.q3xdypn). Cells were counted using NucleoCounter NC-200 and Via1-Cassette and diluted in 0.04% BSA in PBS to a concentration for capture of to attain 8,000-10,000 cells per well according the the 10x Genomics manufacturers protocol.  Cells were loaded into the 10x Chromium Controller according to the manufacturer's protocol at a concentration that would attain between 8,000-10,000 cells/chip position. 10x Genomics VDJ libraries were generated from the 5' 10x Genomics Chromium cDNA libraries as detailed in the manufacturer's protocol.
Experimental Factor: organism part
ascending colon
Experimental Factor: single cell identifier
A38-ACL-1-SC-45N-1
Experimental Factor: cell type
B cell
Experimental Factor: protocol
10x Ig enrichment