Display Settings:

Format

Send to:

Choose Destination
See comment in PubMed Commons below
PLoS One. 2012;7(2):e30794. doi: 10.1371/journal.pone.0030794. Epub 2012 Feb 8.

Highly parallel genome-wide expression analysis of single mammalian cells.

Author information

  • 1Research and Development, Illumina, Inc, San Diego, California, United States of America. jfan@illumina.com

Erratum in

  • PLoS One. 2012;7(2). doi: 10.1371/annotation/3f0ddf41-2e7a-4b56-93c2-2731f2890b19. Loring, Jeanne F [added].

Abstract

BACKGROUND:

We have developed a high-throughput amplification method for generating robust gene expression profiles using single cell or low RNA inputs.

METHODOLOGY/PRINCIPAL FINDINGS:

The method uses tagged priming and template-switching, resulting in the incorporation of universal PCR priming sites at both ends of the synthesized cDNA for global PCR amplification. Coupled with a whole-genome gene expression microarray platform, we routinely obtain expression correlation values of R(2)~0.76-0.80 between individual cells and R(2)~0.69 between 50 pg total RNA replicates. Expression profiles generated from single cells or 50 pg total RNA correlate well with that generated with higher input (1 ng total RNA) (R(2)~0.80). Also, the assay is sufficiently sensitive to detect, in a single cell, approximately 63% of the number of genes detected with 1 ng input, with approximately 97% of the genes detected in the single-cell input also detected in the higher input.

CONCLUSIONS/SIGNIFICANCE:

In summary, our method facilitates whole-genome gene expression profiling in contexts where starting material is extremely limiting, particularly in areas such as the study of progenitor cells in early development and tumor stem cell biology.

PMID:
22347404
[PubMed - indexed for MEDLINE]
PMCID:
PMC3275609
Free PMC Article

Images from this publication.See all images (3)Free text

Figure 1
Figure 2
Figure 3
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Public Library of Science Icon for PubMed Central
    Loading ...
    Write to the Help Desk