Nucleic Acids Research, Vol 25, Issue 10 1999-2004, Copyright © 1997 by Oxford University Press
O Kalinina, I Lebedeva, J Brown and J Silver
We monitored PCR in volumes of the order of 10 nl in glass microcapillaries
using a fluorescence energy transfer assay in which fluorescence increases
if product is made due to template-dependent nucleolytic degradation of an
internally quenched probe (TaqMan assay). This assay detected single
starting template molecules in dilutions of genomic DNA. The results
suggest that it may be feasible to determine the number of template
molecules in a sample by counting the number of positive PCRs in a set of
replicate reactions using terminally diluted sample. Since the assay system
is closed and potentially automatable, it has promise for clinical
applications.
ARTICLES
Nanoliter scale PCR with TaqMan detection
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda MD 20892, USA.
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