Generating High-Quality cDNA Libraries for Efficient Sequencing of T-cell Receptor mRNA

Single-cell omics provide undiluted information about individual cells, such as transcription levels and sequences of highly variable proteins. RNase H-dependent PCR-enabled T cell receptor sequencing (rhTCRseq) allows for an efficient and highly specific analysis of TCR domain heterogeneity for the characterization of the immune response. Miniaturizing the assay helps scale down analysis to the single-cell level without sacrificing sensitivity or data quality.

The key requirement for miniaturization is low-volume liquid handling that can’t be efficiently done manually and demands automation. This application note describes the use of the MANTIS® Liquid Dispenser to construct rhTCRseq cDNA libraries of quality comparable to those made from bulk RNA.
While miniaturization puts strict requirements on liquid handling, this work demonstrates that the MANTIS is well-suited for the miniaturization of moderate-throughput RNAseq and similar -omics applications.

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