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Nat Methods. 2013 Feb;10(2):119-21. doi: 10.1038/nmeth.2305. Epub 2012 Dec 23.

Single-molecule analysis of gene expression using two-color RNA labeling in live yeast.

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1
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York, USA.

Abstract

Live-cell imaging of mRNA yields important insights into gene expression, but it has generally been limited to the labeling of one RNA species and has never been used to count single mRNAs over time in yeast. We demonstrate a two-color imaging system with single-molecule resolution using MS2 and PP7 RNA labeling. We use this methodology to measure intrinsic noise in mRNA levels and RNA polymerase II kinetics at a single gene.

PMID:
23263691
PMCID:
PMC3899799
DOI:
10.1038/nmeth.2305
[Indexed for MEDLINE]
Free PMC Article
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