High-Throughput Single-Cell Labeling (Hi-SCL) for RNA-Seq Using Drop-Based Microfluidics

View/ Open
Author
Shoresh, Noam
Sperling, Ralph A.
Schnall-Levin, Michael
Published Version
https://doi.org/10.1371/journal.pone.0116328Metadata
Show full item recordCitation
Rotem, Assaf, Oren Ram, Noam Shoresh, Ralph A. Sperling, Michael Schnall-Levin, Huidan Zhang, Anindita Basu, Bradley E. Bernstein, and David A. Weitz. 2015. “High-Throughput Single-Cell Labeling (Hi-SCL) for RNA-Seq Using Drop-Based Microfluidics.” PLoS ONE 10 (5): e0116328. doi:10.1371/journal.pone.0116328. http://dx.doi.org/10.1371/journal.pone.0116328.Abstract
The importance of single-cell level data is increasingly appreciated, and significant advances in this direction have been made in recent years. Common to these technologies is the need to physically segregate individual cells into containers, such as wells or chambers of a micro-fluidics chip. High-throughput Single-Cell Labeling (Hi-SCL) in drops is a novel method that uses drop-based libraries of oligonucleotide barcodes to index individual cells in a population. The use of drops as containers, and a microfluidics platform to manipulate them en-masse, yields a highly scalable methodological framework. Once tagged, labeled molecules from different cells may be mixed without losing the cell-of-origin information. Here we demonstrate an application of the method for generating RNA-sequencing data for multiple individual cells within a population. Barcoded oligonucleotides are used to prime cDNA synthesis within drops. Barcoded cDNAs are then combined and subjected to second generation sequencing. The data are deconvoluted based on the barcodes, yielding single-cell mRNA expression data. In a proof-of-concept set of experiments we show that this method yields data comparable to other existing methods, but with unique potential for assaying very large numbers of cells.Other Sources
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4441486/pdf/Terms of Use
This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAACitable link to this page
http://nrs.harvard.edu/urn-3:HUL.InstRepos:16120844
Collections
- FAS Scholarly Articles [17582]
- HMS Scholarly Articles [17782]
Contact administrator regarding this item (to report mistakes or request changes)