Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4155

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Yeast RNA Isolation: Small-Scale

David C. Amberg, Daniel J. Burke, and Jeffrey N. Strathern

This protocol was adapted from "Yeast RNA Isolations," Techniques and Protocols 6, in Methods in Yeast Genetics, 2005 edition, by David C. Amberg, Daniel J. Burke, and Jeffrey N. Strathern. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.

The first 15% of the full text of this article appears below.


INTRODUCTION

This protocol describes rapid, small-scale yeast RNA isolation. It is based on the work of Schmitt et al. (1990). Note that all containers should be washed in RNase Away (Invitrogen) or dry baked for 24 hours at 160°C. All aqueous reagents should be made with 1:1000 volume of DEPC before autoclaving, to inactivate RNases.


MATERIALS

Reagents

recipe AE buffer, chilled

Appropriate yeast culture

caution Dry ice/95% ethanol bath

Ethanol 80% and 100%

caution Phenol equilibrated in AE buffer (pH 5.2)

caution Phenol (pH 5.2):chloroform:isoamyl alcohol (25:24:1)

recipe 10% SDS

recipe Sodium acetate (3 M, pH 5.2)

recipe YPD medium

Equipment

Shaking incubator, preset to an . . . [Full Text of this Article]


METHOD


TROUBLESHOOTING


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