| First Author | Tang F | Year | 2009 |
| Journal | Nat Methods | Volume | 6 |
| Issue | 5 | Pages | 377-82 |
| PubMed ID | 19349980 | Mgi Jnum | J:243335 |
| Mgi Id | MGI:5908166 | Doi | 10.1038/nmeth.1315 |
| Citation | Tang F, et al. (2009) mRNA-Seq whole-transcriptome analysis of a single cell. Nat Methods 6(5):377-82 |
| abstractText | Next-generation sequencing technology is a powerful tool for transcriptome analysis. However, under certain conditions, only a small amount of material is available, which requires more sensitive techniques that can preferably be used at the single-cell level. Here we describe a single-cell digital gene expression profiling assay. Using our mRNA-Seq assay with only a single mouse blastomere, we detected the expression of 75% (5,270) more genes than microarray techniques and identified 1,753 previously unknown splice junctions called by at least 5 reads. Moreover, 8-19% of the genes with multiple known transcript isoforms expressed at least two isoforms in the same blastomere or oocyte, which unambiguously demonstrated the complexity of the transcript variants at whole-genome scale in individual cells. Finally, for Dicer1(-/-) and Ago2(-/-) (Eif2c2(-/-)) oocytes, we found that 1,696 and 1,553 genes, respectively, were abnormally upregulated compared to wild-type controls, with 619 genes in common. |
