Analysis of double-stranded DNA by capillary electrophoresis using poly(ethylene oxide) in the presence of hexadecyltrimethylammonium bromide

Yang-Wei Lin, Huan Tsung Chang

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The impact of hexadecyltrimethylammonium bromide (CTAB) on the separation of ds-DNA by capillary electrophoresis in conjunction with laser-induced fluorescence (CE-LIF) detection using poly(ethylene oxide) (PEO) solution is described. The use of CTAB for improved separation reproducibility and efficiency of DNA has not been demonstrated although it is widely used for controlling the magnitude and direction of electroosmotic flow in CE. With increasing CTAB concentration, the interactions of DNA with ethidium bromide (EtBr) and with the capillary wall decrease. For the separation of DNA fragments with the sizes ranging from several base pairs (bp) to 2176 bp, a polymer solution consisting of 0.75% poly(ethylene oxide), 100 mM TB buffer (pH 8.0), 25 μg/mL EtBr, and 0.36 μg/mL CTAB is proper. Using the PEO solution, we separated a mixture of DNA markers V (pBR 322/HaeIII digest) and VI (pBR 328/BglI digest and pBR 328/HinfI digest) within 8 min at -375 V/cm, with the limit of detection of 2.0 ng/mL based on the peak height for the 18-bp DNA fragment. The method is highly efficient (>106 plate/m), repeatable (RSD of the migration times <1.5%), and sensitive. In addition, it is convenient to fill a capillary (75 μm in diameter) with such a low-viscosity PEO solution by syringe pushing.

Original languageEnglish
Pages (from-to)206-211
Number of pages6
JournalJournal of Chromatography A
Volume1130
Issue number2 SPEC. ISS.
DOIs
Publication statusPublished - 2006 Oct 20

Fingerprint

Ethylene Oxide
Capillary electrophoresis
Capillary Electrophoresis
Polyethylene oxides
Base Pairing
DNA
Ethidium
Electroosmosis
Syringes
Genetic Markers
Viscosity
Limit of Detection
Polymer solutions
Buffers
Polymers
Lasers
Fluorescence
cetrimonium

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

@article{71e7a7740b78474890f6999f94d86fc0,
title = "Analysis of double-stranded DNA by capillary electrophoresis using poly(ethylene oxide) in the presence of hexadecyltrimethylammonium bromide",
abstract = "The impact of hexadecyltrimethylammonium bromide (CTAB) on the separation of ds-DNA by capillary electrophoresis in conjunction with laser-induced fluorescence (CE-LIF) detection using poly(ethylene oxide) (PEO) solution is described. The use of CTAB for improved separation reproducibility and efficiency of DNA has not been demonstrated although it is widely used for controlling the magnitude and direction of electroosmotic flow in CE. With increasing CTAB concentration, the interactions of DNA with ethidium bromide (EtBr) and with the capillary wall decrease. For the separation of DNA fragments with the sizes ranging from several base pairs (bp) to 2176 bp, a polymer solution consisting of 0.75{\%} poly(ethylene oxide), 100 mM TB buffer (pH 8.0), 25 μg/mL EtBr, and 0.36 μg/mL CTAB is proper. Using the PEO solution, we separated a mixture of DNA markers V (pBR 322/HaeIII digest) and VI (pBR 328/BglI digest and pBR 328/HinfI digest) within 8 min at -375 V/cm, with the limit of detection of 2.0 ng/mL based on the peak height for the 18-bp DNA fragment. The method is highly efficient (>106 plate/m), repeatable (RSD of the migration times <1.5{\%}), and sensitive. In addition, it is convenient to fill a capillary (75 μm in diameter) with such a low-viscosity PEO solution by syringe pushing.",
author = "Yang-Wei Lin and Chang, {Huan Tsung}",
year = "2006",
month = "10",
day = "20",
doi = "10.1016/j.chroma.2006.03.050",
language = "English",
volume = "1130",
pages = "206--211",
journal = "Journal of Chromatography",
issn = "0021-9673",
publisher = "Elsevier",
number = "2 SPEC. ISS.",

}

Analysis of double-stranded DNA by capillary electrophoresis using poly(ethylene oxide) in the presence of hexadecyltrimethylammonium bromide. / Lin, Yang-Wei; Chang, Huan Tsung.

In: Journal of Chromatography A, Vol. 1130, No. 2 SPEC. ISS., 20.10.2006, p. 206-211.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Analysis of double-stranded DNA by capillary electrophoresis using poly(ethylene oxide) in the presence of hexadecyltrimethylammonium bromide

AU - Lin, Yang-Wei

AU - Chang, Huan Tsung

PY - 2006/10/20

Y1 - 2006/10/20

N2 - The impact of hexadecyltrimethylammonium bromide (CTAB) on the separation of ds-DNA by capillary electrophoresis in conjunction with laser-induced fluorescence (CE-LIF) detection using poly(ethylene oxide) (PEO) solution is described. The use of CTAB for improved separation reproducibility and efficiency of DNA has not been demonstrated although it is widely used for controlling the magnitude and direction of electroosmotic flow in CE. With increasing CTAB concentration, the interactions of DNA with ethidium bromide (EtBr) and with the capillary wall decrease. For the separation of DNA fragments with the sizes ranging from several base pairs (bp) to 2176 bp, a polymer solution consisting of 0.75% poly(ethylene oxide), 100 mM TB buffer (pH 8.0), 25 μg/mL EtBr, and 0.36 μg/mL CTAB is proper. Using the PEO solution, we separated a mixture of DNA markers V (pBR 322/HaeIII digest) and VI (pBR 328/BglI digest and pBR 328/HinfI digest) within 8 min at -375 V/cm, with the limit of detection of 2.0 ng/mL based on the peak height for the 18-bp DNA fragment. The method is highly efficient (>106 plate/m), repeatable (RSD of the migration times <1.5%), and sensitive. In addition, it is convenient to fill a capillary (75 μm in diameter) with such a low-viscosity PEO solution by syringe pushing.

AB - The impact of hexadecyltrimethylammonium bromide (CTAB) on the separation of ds-DNA by capillary electrophoresis in conjunction with laser-induced fluorescence (CE-LIF) detection using poly(ethylene oxide) (PEO) solution is described. The use of CTAB for improved separation reproducibility and efficiency of DNA has not been demonstrated although it is widely used for controlling the magnitude and direction of electroosmotic flow in CE. With increasing CTAB concentration, the interactions of DNA with ethidium bromide (EtBr) and with the capillary wall decrease. For the separation of DNA fragments with the sizes ranging from several base pairs (bp) to 2176 bp, a polymer solution consisting of 0.75% poly(ethylene oxide), 100 mM TB buffer (pH 8.0), 25 μg/mL EtBr, and 0.36 μg/mL CTAB is proper. Using the PEO solution, we separated a mixture of DNA markers V (pBR 322/HaeIII digest) and VI (pBR 328/BglI digest and pBR 328/HinfI digest) within 8 min at -375 V/cm, with the limit of detection of 2.0 ng/mL based on the peak height for the 18-bp DNA fragment. The method is highly efficient (>106 plate/m), repeatable (RSD of the migration times <1.5%), and sensitive. In addition, it is convenient to fill a capillary (75 μm in diameter) with such a low-viscosity PEO solution by syringe pushing.

UR - http://www.scopus.com/inward/record.url?scp=33748967508&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748967508&partnerID=8YFLogxK

U2 - 10.1016/j.chroma.2006.03.050

DO - 10.1016/j.chroma.2006.03.050

M3 - Article

C2 - 16597443

AN - SCOPUS:33748967508

VL - 1130

SP - 206

EP - 211

JO - Journal of Chromatography

JF - Journal of Chromatography

SN - 0021-9673

IS - 2 SPEC. ISS.

ER -