Resolution of Budding Yeast Chromosomes Using Pulsed-Field Gel Electrophoresis
- 17 July 2013
- book chapter
- Published by Springer Nature
- Vol. 1054, 195-207
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.Keywords
This publication has 14 references indexed in Scilit:
- Chromosome length influences replication-induced topological stressNature, 2011
- Trf4 targets ncRNAs from telomeric and rDNA spacer regions and functions in rDNA copy number controlThe EMBO Journal, 2007
- Recombination Regulation by Transcription-Induced Cohesin Dissociation in rDNA RepeatsScience, 2005
- Methyl methanesulfonate (MMS) produces heat-labile DNA damage but no detectable in vivo DNA double-strand breaksNucleic Acids Research, 2005
- A Role forSaccharomyces cerevisiaeChk1p in the Response to Replication BlocksMolecular Biology of the Cell, 2004
- NuA4 Subunit Yng2 Function in Intra-S-Phase DNA Damage ResponseMolecular and Cellular Biology, 2002
- Pulsed‐field gel analysis of the pattern of DNA double‐strand breaks in the Saccharomyces genome during meiosisDevelopmental Genetics, 1992
- Separation of Large DNA Molecules by Contour-Clamped Homogeneous Electric FieldsScience, 1986
- Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresisNucleic Acids Research, 1984
- Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresisCell, 1984