We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Highly Parallel Genome-Wide Expression Analysis of Single Mammalian Cells.
- Authors
Jian-Bing Fan; Jing Chen; April, Craig S.; Fisher, Jeffrey S.; Klotzle, Brandy; Bibikova, Marina; Kaper, Fiona; Ronaghi, Mostafa; Linnarsson, Sten; Ota, Takayo; Jeremy Chien; Laurent, Louise C.; Nisperos, Sean V.; Chen, Gina Y.; Zhong, Jiang F.
- 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 wholegenome gene expression microarray platform, we routinely obtain expression correlation values of R2∼0.76-0.80 between individual cells and R2∼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) (R2∼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.
- Publication
PLoS ONE, 2012, Vol 7, Issue 2, p1
- ISSN
1932-6203
- Publication type
Academic Journal
- DOI
10.1371/journal.pone.0030794