Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate.

Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate. Research Abstract Details 

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Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate. Abstract Text:

Jifeng Chen,Musundi Wabuyele,Hengwu Chen,Donald Patterson,Mateusz Hupert,Hamed Shadpour,Dimitris Nikitopoulos,Steven A Soper,

This paper presents a novel method for DNA thermal amplification using the polymerase chain reaction (PCR) in an electrokinetically driven synchronized continuous flow PCR (EDS-CF-PCR) configuration carried out in a microfabricated polycarbonate (PC) chip. The synchronized format allowed patterning a shorter length microchannel for the PCR compared to nonsynchronized continuous flow formats, permitting the use of smaller applied voltages when the flow is driven electrically and also allowed flexibility in selecting the cycle number without having to change the microchip architecture. A home-built temperature control system was developed to precisely configure three isothermal zones on the chip for denaturing (95 degrees C), annealing (55 degrees C), and extension (72 degrees C) within a single-loop channel. DNA templates were introduced into the PCR reactor, which was filled with the PCR cocktail, by electrokinetic injection. The PCR cocktail consisted of low salt concentrations (KCl) to reduce the current in the EDS-CF-PCR device during cycling. To control the EOF in the PC microchannel to minimize dilution effects as the DNA "plug" was shuttled through the temperature zones, Polybrene was used as a dynamic coating, which resulted in reversal of the EOF. The products generated from 15, 27, 35, and 40 EDS-CF-PCR amplification cycles were collected and analyzed using microchip electrophoresis with LIF detection for fragment sizing. The results showed that the EDS-CF-PCR format produced results similar to that of a conventional block thermal cycler with leveling effects observed for amplicon generation after approximately 25 cycles. To the best of our knowledge, this is the first report of electrokinetically driven synchronized PCR performed on chip.

Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate. Publishing Authors By Initials

J Chen,M Wabuyele,H Chen,D Patterson,M Hupert,H Shadpour,D Nikitopoulos,SA Soper,

For similar investigative techniques: genetic techniques: nucleic acid amplification techniques: polymerase chain reaction research abstracts see: investigative techniques: genetic techniques: nucleic acid amplification techniques: polymerase chain reaction research

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Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Analytical chemistry

VOLUME: 77

Page Numbers: 658-66

Journal Abbreviation: Anal. Chem.

ISSN: 0003-2700

DAY: 15

MONTH: Jan

YEAR: 2005

Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 370536

Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate. Keywords Mesh Terms:

KEYWORDS: Polymerase Chain Reaction

MESH TERMS: methods

Chemical & Substance for Abstract: Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate. Information

Substance Name: DNA

Registry Number: 9007-49-2

Grant and Affiliation Information for Electrokinetically synchronized polymerase chain reaction microchip fabricated in polycarbonate.

AFFILIATION: Department of Chemistry and Center for BioModular Multi-Scale Systems, and Department of Mechanical Engineering, Louisiana State University, Baton Rouge, Louisiana 70803, USA.

Country: United States

AGENCY: United States NIBIB

GRANT: R24-EB002115

ACRONYM: EB

MEDLINETA: Anal Chem

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