Improving the length-fractionation of DNA during capillary electrophoresis

Gary A. Griess, Philip Serwer

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

The present study develops a path-lengthening strategy for capillary electrophoresis of short double-stranded DNA molecules, in an aqueous solution of neutral polymer (hydroxypropylmethylcellulose). Tests of the dependence of fractionations on pulse times reveal the operation of at least one mechanism in addition to increase in effective path length. Electrophoresis is performed in the following two-stage cycles (cyclic electrophoresis): The first analysis-stage of each cycle is a constant field (forward) capillary electrophoresis. This analysis-stage reveals the length distribution of the shortest DNA molecules not previously analyzed. The second, enhancement-stage of each cycle is zero-integrated field electrophoresis (ZIFE). The enhancement-stage improves the DNA length-fractionation for the next DNA molecules to be analyzed. A slight reverse migration occurs in the enhancement-stage. Increase in both peak separation and peak sharpness contribute to improvement in the length-fractionation of DNA molecules.

Original languageEnglish (US)
Pages (from-to)4320-4327
Number of pages8
JournalELECTROPHORESIS
Volume22
Issue number20
DOIs
Publication statusPublished - Dec 1 2001

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Keywords

  • Capillary electrophoresis
  • Cellulose derivative
  • Nongelled polymer sieving
  • Pulsed-field electrophoresis
  • Selectivity enhancement

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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