| Autor: |
Fuchou Tang, Barbacioru, Catalin, Yangzhou Wang, Nordman, Ellen, Clarence Lee, Nanlan Xu, Xiaohui Wang, Bodeau, John, Tuch, Brian B, Siddiqui, Asim, Lao, Kaiqin, Surani, M. Azim |
| Předmět: |
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| Zdroj: |
Nature Methods; May2009, Vol. 6 Issue 5, p377-382, 6p, 1 Diagram, 1 Chart, 4 Graphs |
| Abstrakt: |
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. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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