Stabilization and electrophoretic analysis of meiotic recombination intermediates in Saccharomyces cerevisiae.
Joint Molecule (JM) recombination intermediates result from DNA strand-exchange between homologous chromosomes. Physical monitoring of JM formation in budding yeast has provided a wealth of information about the timing and mechanism of meiotic recombination. These assays are especially informative when applied to the analysis of mutants for which genetic analysis of recombination is impossible, i.e. mutants that die during meiosis. This chapter describes three distinct methods to stabilize JMs against thermally driven dissolution as well as electrophoretic approaches to resolve and detect JMs at two well-characterized recombination hotspots.
Oh
Steve D
SD
Department of Microbiology, University of California, Davis, CA, USA.
Jessop
Lea
L
Lao
Jessica P
JP
Allers
Thorsten
T
Lichten
Michael
M
Hunter
Neil
N
D000465:Algorithms; D043171:Chromosomal Instability / Q000502:physiology*; D004271:DNA, Fungal / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel / Q000379:methods; D008540:Meiosis / Q000235:genetics*; D008954:Models, Biological; D011995:Recombination, Genetic / Q000235:genetics*; D015183:Restriction Mapping / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics*
D004271:DNA, Fungal