Unlimited increase in the resolution of DNA ladders

Gary A. Griess, Eric Rogers, Philip Serwer

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Fractionation of DNA ladders by gel electrophoresis is limited by the progressive compressing of the long DNA end of a ladder. Improvement in the resolution of this DNA is achieved by use of the following two-step electrophoresis. Initially, the DNA ladder is fractionated by conventional constant field agarose gel electrophoresis. Subsequently, gel electrophoresis is performed in the reverse direction by pulsing the electrical field (PFGE). A newly developed type of pulsing is used, which causes inversion of a double-stranded DNA ladder: the distance migrated increases as the length of the DNA molecule increases. Thus, the resolution of DNA bands continues to increase during the PFGE. These two stages of electrophoresis are serially repeated. Eventually, both the short and the long DNA ends of the ladder migrate out of the gel while a selected region of the ladder undergoes progressive increase in resolution during back-and-forth migration. Improved resolution of DNA bands is achieved, without a known limit.

Original languageEnglish (US)
Pages (from-to)859-864
Number of pages6
JournalELECTROPHORESIS
Volume21
Issue number5
DOIs
StatePublished - 2000

Keywords

  • Agarose gel
  • DNA fractionation
  • Field effects
  • Pulsed field electrophoresis

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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