Resolution of budding yeast chromosomes using pulsed-field gel electrophoresis

Aziz El Hage, Jonathan Houseley

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract

Pulsed-field gel electrophoresis (PFGE) is a technique that resolves chromosome-sized DNA molecules in an agarose gel. As well as DNA mapping and karyotyping applications, PFGE techniques are well adapted to follow DNA rearrangements over time in a quantitative manner. Because of the very large sizes of the DNA molecules analyzed, DNA preparation, electrophoresis, and Southern blotting processes present unique challenges in PFGE experiments. In this chapter, we describe a robust PFGE protocol covering the preparation of intact Saccharomyces cerevisiae chromosomal DNA, specific running conditions for the resolution of small, medium- and large-sized chromosomes and their by-products, and basic Southern blotting and hybridization instructions for the analysis of these molecules.
Original languageEnglish
Title of host publicationDNA electrophoresis
EditorsSvetlana Makovets
Place of PublicationUSA
PublisherSpringer Nature
Pages195-207
Number of pages13
Volume1054
ISBN (Electronic)978-1-62703-565-1
ISBN (Print)978-1-62703-564-4
DOIs
Publication statusPublished - 17 Jul 2013

Publication series

NameMethods in Molecular Biology

Keywords

  • Saccharomyces cerevisiae
  • Pulsed-field gel electrophoresis (PFGE)
  • Contour-clamped homogeneous electric field (CHEF)
  • Agarose embedded yeast DNA
  • Southern blot
  • Hybridization

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