Comparative analysis of single-cell RNA sequencing platforms and methods

Single-cell RNA sequencing (scRNA-seq) offers great new opportunities for increasing our understanding of complex biological processes. In particular, development of an accurate Human Cell Atlas is largely dependent on the rapidly advancing technologies and molecular chemistries employed in scRNA-seq. These advances have already allowed an increase in throughput for scRNA-seq from 96 to 80,000 cells on a single instrument run by capturing cells within nano-liter droplets. While this increase in throughput is critical for many experimental questions, a thorough comparison between microfluidic-, plate-, and droplet-based technologies or between multiple available platforms utilizing these technologies is largely lacking.

Here, a team led by researchers at the University of Rochester Medical Center report scRNA-seq data from SUM149PT cells treated with the histone deacetylase inhibitor TSA vs. untreated controls across several scRNA-seq platforms (Fluidigm C1, WaferGen iCell8, 10X Genomics Chromium Controller, and Illumina/BioRad ddSEQ). The primary goal of this project was to demonstrate RNA sequencing (RNA-seq) methods for profiling the ultra-low amounts of RNA present in individual cells, and this report discusses the results of the study as well as technical challenges/lesson learned and present general guidelines for best practices in sample preparation and analysis.

Experimental setup

rna-seq

Cells from the sum149PT breast cancer cell line were subjected to DMSO and TSA treatment and subsequently sequenced using different technologies.

Ashton JM, Rehrauer H, Myers J, Myers J, Zanche M, Balys M, Foox J, Mason CE, Steen R, Kuentzel M, Aquino C, Garcia-Reyero N, Chittur SV. (2020) Comparative analysis of single-cell RNA Sequencing Platforms and Methods. bioRXiv [online preprint]. [abstract]

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