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Rapid DNA sequencing of more than 1000 bases per run by capillary electrophoresis using replaceable linear polyacrylamide solutions.

Publication ,  Journal Article
Carrilho, E; Ruiz-Martinez, MC; Berka, J; Smirnov, I; Goetzinger, W; Miller, AW; Brady, D; Karger, BL
Published in: Analytical chemistry
October 1996

The read length for DNA sequencing using capillary electrophoresis and replaceable linear polyacrylamide (LPA) solutions has been extended to more than 1000 bases with a run time of 80 min. This result was successfully achieved through the combined use of cycle sequencing with dye-labeled primers, improved matrix and separation conditions, and enhanced base-calling software. The influences of LPA molecular weight and concentration on separation were investigated. Additionally, the separation buffer, column temperature, and electric field were adjusted to increase the number of resolvable DNA fragments per run while maintaining an enhanced separation speed. Using low concentrations [2% (w/v)] of high molecular weight LPA polymers (> 5.5 x 10(6) Da), elevated column temperature (50 degrees C) and moderately high field (150 V/cm), rapid sequencing analysis for more than 1000 bases on a model ssM13mp18 template was obtained with 96.8% accuracy.

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Published In

Analytical chemistry

DOI

EISSN

1520-6882

ISSN

0003-2700

Publication Date

October 1996

Volume

68

Issue

19

Start / End Page

3305 / 3313

Related Subject Headings

  • Sequence Analysis, DNA
  • Molecular Weight
  • Molecular Sequence Data
  • Electrophoresis, Capillary
  • DNA
  • Base Sequence
  • Base Composition
  • Analytical Chemistry
  • Acrylic Resins
  • 4004 Chemical engineering
 

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Carrilho, E., Ruiz-Martinez, M. C., Berka, J., Smirnov, I., Goetzinger, W., Miller, A. W., … Karger, B. L. (1996). Rapid DNA sequencing of more than 1000 bases per run by capillary electrophoresis using replaceable linear polyacrylamide solutions. Analytical Chemistry, 68(19), 3305–3313. https://doi.org/10.1021/ac960411r
Carrilho, E., M. C. Ruiz-Martinez, J. Berka, I. Smirnov, W. Goetzinger, A. W. Miller, D. Brady, and B. L. Karger. “Rapid DNA sequencing of more than 1000 bases per run by capillary electrophoresis using replaceable linear polyacrylamide solutions.Analytical Chemistry 68, no. 19 (October 1996): 3305–13. https://doi.org/10.1021/ac960411r.
Carrilho E, Ruiz-Martinez MC, Berka J, Smirnov I, Goetzinger W, Miller AW, et al. Rapid DNA sequencing of more than 1000 bases per run by capillary electrophoresis using replaceable linear polyacrylamide solutions. Analytical chemistry. 1996 Oct;68(19):3305–13.
Carrilho, E., et al. “Rapid DNA sequencing of more than 1000 bases per run by capillary electrophoresis using replaceable linear polyacrylamide solutions.Analytical Chemistry, vol. 68, no. 19, Oct. 1996, pp. 3305–13. Epmc, doi:10.1021/ac960411r.
Carrilho E, Ruiz-Martinez MC, Berka J, Smirnov I, Goetzinger W, Miller AW, Brady D, Karger BL. Rapid DNA sequencing of more than 1000 bases per run by capillary electrophoresis using replaceable linear polyacrylamide solutions. Analytical chemistry. 1996 Oct;68(19):3305–3313.
Journal cover image

Published In

Analytical chemistry

DOI

EISSN

1520-6882

ISSN

0003-2700

Publication Date

October 1996

Volume

68

Issue

19

Start / End Page

3305 / 3313

Related Subject Headings

  • Sequence Analysis, DNA
  • Molecular Weight
  • Molecular Sequence Data
  • Electrophoresis, Capillary
  • DNA
  • Base Sequence
  • Base Composition
  • Analytical Chemistry
  • Acrylic Resins
  • 4004 Chemical engineering