GSM5668737: DS799Feature Barcoding; Homo sapiens; OTHER
10X was used for all single cell experiements following the 5' v1.1 immune profiling with feature barcoding protocol. For the bulk TCR, CD4+ T cells were isolated by negative selection, and total RNA was extracted using the Zymo Directzol RNA kit. 10X Immune Profiling V1.1. Briefly, after cDNA amplification, cDNA is separated into high molecular weight (HMW) and low molecular weight (LMW) fractions. The LMW fraction is library prepped to generate CITE-seq libraries of surface protein expression. The HMW is sheered enzymatically, end repaired, and adapters are ligated on to generate the transcriptome library. In parallel, targetted amplification of TCR sequences occurs on the HMW fraction.