Membrane-mediated ultrafast restriction digestion and subsequent rapid gel microchip electrophoresis of DNA

A. Guttman, Z. Rónai, C. Barta, Yu Ming Hou, M. Sasvári, Xun Wang, Steven P. Briggs

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Ultrafast, membrane-mediated restriction digestion of DNA molecules followed by rapid gel microchip electrophoresis of the resulting fragments is described. Combination of restriction endonuclease digestion on small pore-size microfibrous membranes with sample loading and electrophoresis analysis in a multilane (up to 96) format resulted in very fast restriction digest based microscale DNA analysis. Complete digestion of several nanogram target DNA was accomplished on the microporous membrane at room temperature just in a few minutes with a single or a combination of various restriction enzymes, using only submicroliter quantities of samples and reagents. The reaction mixture containing membrane also served as sample loading device for the subsequent gel microchip electrophoresis based analysis. This work establishes methods for high-speed, high-throughput DNA analysis, featuring extremely low sample and reagent consumption, and fast restriction digestion in combination with sample loading and rapid gel microchip analysis of the resulting fragments. The entire restriction digestion, sample loading and electrophoresis analysis process required less than 20 min.

Original languageEnglish
Pages (from-to)1524-1530
Number of pages7
JournalElectrophoresis
Volume23
Issue number10
DOIs
Publication statusPublished - 2002

Fingerprint

Microchip Electrophoresis
Electrophoresis
Digestion
Gels
Membranes
DNA
Microchip Analytical Procedures
DNA Restriction Enzymes
Pore size
Throughput
Molecules
Enzymes
Equipment and Supplies
Temperature

Keywords

  • DNA analysis
  • Microchip electrophoresis
  • Restriction digestion

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Membrane-mediated ultrafast restriction digestion and subsequent rapid gel microchip electrophoresis of DNA. / Guttman, A.; Rónai, Z.; Barta, C.; Hou, Yu Ming; Sasvári, M.; Wang, Xun; Briggs, Steven P.

In: Electrophoresis, Vol. 23, No. 10, 2002, p. 1524-1530.

Research output: Contribution to journalArticle

@article{4260d0c501ee4aafabef7374caae867b,
title = "Membrane-mediated ultrafast restriction digestion and subsequent rapid gel microchip electrophoresis of DNA",
abstract = "Ultrafast, membrane-mediated restriction digestion of DNA molecules followed by rapid gel microchip electrophoresis of the resulting fragments is described. Combination of restriction endonuclease digestion on small pore-size microfibrous membranes with sample loading and electrophoresis analysis in a multilane (up to 96) format resulted in very fast restriction digest based microscale DNA analysis. Complete digestion of several nanogram target DNA was accomplished on the microporous membrane at room temperature just in a few minutes with a single or a combination of various restriction enzymes, using only submicroliter quantities of samples and reagents. The reaction mixture containing membrane also served as sample loading device for the subsequent gel microchip electrophoresis based analysis. This work establishes methods for high-speed, high-throughput DNA analysis, featuring extremely low sample and reagent consumption, and fast restriction digestion in combination with sample loading and rapid gel microchip analysis of the resulting fragments. The entire restriction digestion, sample loading and electrophoresis analysis process required less than 20 min.",
keywords = "DNA analysis, Microchip electrophoresis, Restriction digestion",
author = "A. Guttman and Z. R{\'o}nai and C. Barta and Hou, {Yu Ming} and M. Sasv{\'a}ri and Xun Wang and Briggs, {Steven P.}",
year = "2002",
doi = "10.1002/1522-2683(200205)23:10<1524::AID-ELPS1524>3.0.CO;2-Y",
language = "English",
volume = "23",
pages = "1524--1530",
journal = "Electrophoresis",
issn = "0173-0835",
publisher = "Wiley-VCH Verlag",
number = "10",

}

TY - JOUR

T1 - Membrane-mediated ultrafast restriction digestion and subsequent rapid gel microchip electrophoresis of DNA

AU - Guttman, A.

AU - Rónai, Z.

AU - Barta, C.

AU - Hou, Yu Ming

AU - Sasvári, M.

AU - Wang, Xun

AU - Briggs, Steven P.

PY - 2002

Y1 - 2002

N2 - Ultrafast, membrane-mediated restriction digestion of DNA molecules followed by rapid gel microchip electrophoresis of the resulting fragments is described. Combination of restriction endonuclease digestion on small pore-size microfibrous membranes with sample loading and electrophoresis analysis in a multilane (up to 96) format resulted in very fast restriction digest based microscale DNA analysis. Complete digestion of several nanogram target DNA was accomplished on the microporous membrane at room temperature just in a few minutes with a single or a combination of various restriction enzymes, using only submicroliter quantities of samples and reagents. The reaction mixture containing membrane also served as sample loading device for the subsequent gel microchip electrophoresis based analysis. This work establishes methods for high-speed, high-throughput DNA analysis, featuring extremely low sample and reagent consumption, and fast restriction digestion in combination with sample loading and rapid gel microchip analysis of the resulting fragments. The entire restriction digestion, sample loading and electrophoresis analysis process required less than 20 min.

AB - Ultrafast, membrane-mediated restriction digestion of DNA molecules followed by rapid gel microchip electrophoresis of the resulting fragments is described. Combination of restriction endonuclease digestion on small pore-size microfibrous membranes with sample loading and electrophoresis analysis in a multilane (up to 96) format resulted in very fast restriction digest based microscale DNA analysis. Complete digestion of several nanogram target DNA was accomplished on the microporous membrane at room temperature just in a few minutes with a single or a combination of various restriction enzymes, using only submicroliter quantities of samples and reagents. The reaction mixture containing membrane also served as sample loading device for the subsequent gel microchip electrophoresis based analysis. This work establishes methods for high-speed, high-throughput DNA analysis, featuring extremely low sample and reagent consumption, and fast restriction digestion in combination with sample loading and rapid gel microchip analysis of the resulting fragments. The entire restriction digestion, sample loading and electrophoresis analysis process required less than 20 min.

KW - DNA analysis

KW - Microchip electrophoresis

KW - Restriction digestion

UR - http://www.scopus.com/inward/record.url?scp=0036072740&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036072740&partnerID=8YFLogxK

U2 - 10.1002/1522-2683(200205)23:10<1524::AID-ELPS1524>3.0.CO;2-Y

DO - 10.1002/1522-2683(200205)23:10<1524::AID-ELPS1524>3.0.CO;2-Y

M3 - Article

C2 - 12116164

AN - SCOPUS:0036072740

VL - 23

SP - 1524

EP - 1530

JO - Electrophoresis

JF - Electrophoresis

SN - 0173-0835

IS - 10

ER -