Lighting Up: Analysis Of Yeast RAS Genes Using A Chemiluminescent Southern Blot Procedure

 

Karen A. Malatesta and Linda B. Cholewiak

Tested Studies in Laboratory Teaching, 1997, Volume 18

Abstract

Genomic and plasmid-encoded RAS2 genes from Baker’s and brewing yeast strains will be analyzed using a shortened Southern blot procedure employing chemiluminescence. Unique parts of this procedure are the gravity-driven “upside down” transfer and a rotisserie-like hybridization oven, which greatly facilitate shortening the transfer time to 45 minutes and the hybridization step to 60 minutes. Chemiluminescence obviates problems associated with radioisotopes without sacrificing sensitivity: the plasmid-borne RAS gene can be visualized within 15 seconds and the single-copy yeast genomic RAS2 gene, within a minute. Topics for discussion related to this laboratory can include the concepts of RFLPs, evolutionary conservation of DNA sequences and the genetic basis of cancer.

Keywords:  agarose gel electrophoresis, yeast, southern blot, chemiluminescence, RAS

Boston University (1996)