Document Detail


High-throughput droplet PCR.
MedLine Citation:
PMID:  20117212     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The polymerase chain reaction has facilitated the ready analysis of nucleic acids. A next challenge requires the development of means to unravel the complexity of heterogeneous tissues. This has presented the task of producing massively parallelized quantitative nucleic acid data from the cellular constituents of tissues. The production of aqueous droplets in a two phase flow is shown to be readily and routinely facilitated by miniaturized fluidic devices. Droplets serve as ideal means to package a future generation of PCR, offering an enhanced handling potential by virtue of reactant containment, to concurrently eliminate both contamination and sample loss. This containment also enables the measurement of nucleic acids from populations of cells, or molecules by means of high throughput, single cell analysis. Details are provided for the production of a prototype micro-fluidic device which shows the production and stable flow of droplets which we suggest will be suitable for droplet-based continuous flow micro-fluidic PCR. Suggestions are also made as to the optimal fabrication techniques and the importance of device calibration.
Authors:
Amelia L Markey; Stephan Mohr; Philip J R Day
Publication Detail:
Type:  Journal Article     Date:  2010-02-02
Journal Detail:
Title:  Methods (San Diego, Calif.)     Volume:  50     ISSN:  1095-9130     ISO Abbreviation:  Methods     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-03-10     Completed Date:  2010-05-28     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9426302     Medline TA:  Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  277-81     Citation Subset:  IM    
Copyright Information:
Copyright 2010 Elsevier Inc. All rights reserved.
Affiliation:
School of Chemical Engineering and Analytical Sciences, Manchester Interdisciplinary Biocentre, University of Manchester, UK.
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MeSH Terms
Descriptor/Qualifier:
Calibration
Microfluidic Analytical Techniques / instrumentation*,  methods
Polymerase Chain Reaction / instrumentation,  methods*
Temperature

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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