Highly Multiplexed, Single Cell Transcriptomic Analysis of T-Cells by Microfluidic PCR
- PMID: 27787802
- DOI: 10.1007/978-1-4939-6548-9_15
Highly Multiplexed, Single Cell Transcriptomic Analysis of T-Cells by Microfluidic PCR
Abstract
Recently, technologies have been developed to measure expression of 96 (or more) mRNA transcripts at once from a single cell. Here we describe methods and important considerations for use of Fluidigm's BioMark platform for multiplexed single cell gene expression. We describe how to qualify primer/probes, select genes to examine in 96-parameter panels, perform the reverse transcription/cDNA synthesis step, and operate the instrument. In addition, we describe data analysis considerations. This technology has enormous value for characterizing the heterogeneity of T-cells, thereby providing a useful tool for immune monitoring.
Keywords: Microfluidic; Quantitative competitive real-time PCR; Single cell analysis; T-cell; Transcriptome.
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