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

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Author:

Karen Malatesta, Linda B. Cholewiak

Subject:

Science and Technology

Institution Name:

Princeton University

Collection:

Association for Biology Laboratory Education (ABLE)

Grade Level:

Post-secondary

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.

Course Type:

Learning Module

Languages:

English

Material Type:

Activities and Labs, Lesson Plans

Media Format:

Downloadable docs

Conditions of Use:

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All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without the prior written permission of the copyright owner. Use solely at one's own institution with no intent for profit is excluded from the preceding copyright restriction, unless otherwise noted on the copyright notice of the individual chapter in this volume. Proper credit to this publication must be included in your laboratory outline for each use; a sample citation is given [on the copyright page of each volume]. Upon obtaining permission or with the "sole use at one's own institution" exclusion, ABLE strongly encourages individuals to use the exercises in this proceedings volume in their teaching program.