alias: S3
title: RNA-seq of T cell receptors: adult male 34y pbmc
design: T cell receptor  and  cDNA was captured in emulsion at a single cell level. Specific reverse transcription primers were used to target T cell receptor  and  chains along with the addition of unique molecular identifiers to account for PCR bias. The resulting cDNAs were then subjected to emulsion PCR, during which they anneal to amplified droplet barcode strands resulting in a DNA product containing droplet barcode (13-18 nucleotides), a unique molecular identifier (UMI) and a target sequence. After emulsion RT and PCR, the emulsion was broken and the products were purified and subjected to target enrichment PCR, where primers specific to constant regions of T cell receptors and to the droplet barcode constant sequence were used. Sequencing libraries from each emulsion were uniquely tagged with Illumina index sequences for multiplexed next-generation sequencing. The full-length TCR variable cDNA was then sequenced on Illumina MiSeq (paired-end 325+300 bp).
identifiers.namespace: SUB4930779
identifiers.id: S3