Anal Chem. 2009 Jun 15;81(12):4813-21.

Droplet-based microfluidic systems for high-throughput single DNA molecule isothermal amplification and analysis.

Mazutis L, Araghi AF, Miller OJ, Baret JC, Frenz L, Janoshazi A, Taly V, Miller BJ, Hutchison JB, Link D, Griffiths AD, Ryckelynck M.

Source

Institut de Science et d'Ingenierie Supramoleculaire, Universite de Strasbourg, CNRS UMR 7006, 8 allee Gaspard Monge, 67083 Strasbourg Cedex, France.

Abstract

We have developed a method for high-throughput isothermal amplification of single DNA molecules in a droplet-based microfluidic system. DNA amplification in droplets was analyzed using an intercalating fluorochrome, allowing fast and accurate "digital" quantification of the template DNA based on the Poisson distribution of DNA molecules in droplets. The clonal amplified DNA in each 2 pL droplet was further analyzed by measuring the enzymatic activity of the encoded proteins after fusion with a 15 pL droplet containing an in vitro translation system.

PMID:: 19518143; [PubMed - indexed for MEDLINE]

Publication Types, MeSH Terms, Substances, Grant Support

Publication Types

Research Support, Non-U.S. Gov't

MeSH Terms

Substances

Grant Support

Medical Research Council/United Kingdom

LinkOut - more resources

Full Text Sources

Supplemental Content

Related citations

Review Microfluidic DNA amplification--a review. [Anal Chim Acta. 2009]
Review Microfluidic DNA amplification--a review.
Zhang Y, Ozdemir P. Anal Chim Acta. 2009 Apr 13; 638(2):115-25. Epub 2009 Mar 4.
Continuous-flow polymerase chain reaction of single-copy DNA in microfluidic microdroplets. [Anal Chem. 2009]
Continuous-flow polymerase chain reaction of single-copy DNA in microfluidic microdroplets.
Schaerli Y, Wootton RC, Robinson T, Stein V, Dunsby C, Neil MA, French PM, Demello AJ, Abell C, Hollfelder F. Anal Chem. 2009 Jan 1; 81(1):302-6.
Multi-step microfluidic droplet processing: kinetic analysis of an in vitro translated enzyme. [Lab Chip. 2009]
Multi-step microfluidic droplet processing: kinetic analysis of an in vitro translated enzyme.
Mazutis L, Baret JC, Treacy P, Skhiri Y, Araghi AF, Ryckelynck M, Taly V, Griffiths AD. Lab Chip. 2009 Oct 21; 9(20):2902-8. Epub 2009 Aug 6.
Loop-mediated isothermal amplification integrated on microfluidic chips for point-of-care quantitative detection of pathogens. [Anal Chem. 2010]
Loop-mediated isothermal amplification integrated on microfluidic chips for point-of-care quantitative detection of pathogens.
Fang X, Liu Y, Kong J, Jiang X. Anal Chem. 2010 Apr 1; 82(7):3002-6.
Review Microsystems technology and biosensing. [Adv Biochem Eng Biotechnol. 2008]
Review Microsystems technology and biosensing.
Sathuluri RR, Yamamura S, Tamiya E. Adv Biochem Eng Biotechnol. 2008; 109:285-350.

See reviews... See all...

Cited by 4 PubMed Central articles

System Integration - A Major Step toward Lab on a Chip. [J Biol Eng. 2011]
System Integration - A Major Step toward Lab on a Chip.
Sin ML, Gao J, Liao JC, Wong PK. J Biol Eng. 2011 May 25; 5:6. Epub 2011 May 25.
Microfluidic compartmentalized directed evolution. [Chem Biol. 2010]
Microfluidic compartmentalized directed evolution.
Paegel BM, Joyce GF. Chem Biol. 2010 Jul 30; 17(7):717-24.
Digital PCR on a SlipChip. [Lab Chip. 2010]
Digital PCR on a SlipChip.
Shen F, Du W, Kreutz JE, Fok A, Ismagilov RF. Lab Chip. 2010 Oct 21; 10(20):2666-72. Epub 2010 Jul 1.

See all...

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
Compound (MeSH Keyword)
PubChem chemical compound records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Substance (MeSH Keyword)
PubChem chemical substance (submitted) records that are classified under the same Medical Subject Headings (MeSH) controlled vocabulary as the current articles.
Cited in PMC
Full-text articles in the PubMed Central Database that cite the current articles.

Recent activity

Clear Turn Off Turn On

Droplet-based microfluidic systems for high-throughput single DNA molecule isoth...
Droplet-based microfluidic systems for high-throughput single DNA molecule isothermal amplification and analysis.
Anal Chem. 2009 Jun 15 ;81(12):4813-21.

PubMed
High-efficiency gene transfection of macrophages by lipoplexes.
High-efficiency gene transfection of macrophages by lipoplexes.
Int J Pharm. 2000 Sep 25 ;206(1-2):97-104.

PubMed
Benign lymphangioendothelioma manifested clinically as actinic keratosis.
Benign lymphangioendothelioma manifested clinically as actinic keratosis.
Cutis. 2001 Jan ;67(1):29-30.

PubMed
Reinforcement of anchorage during orthodontic brace treatment with implants or o...
Reinforcement of anchorage during orthodontic brace treatment with implants or other surgical methods.
Cochrane Database Syst Rev. 2007 Jul 18 ;(3):CD005098.

PubMed
Emergency: pediatric orthopnea and total airway obstruction.
Emergency: pediatric orthopnea and total airway obstruction.
Am J Nurs. 2007 Apr ;107(4):40-3.

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Display Settings:

Send to: