Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot3298

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Growth of S. cerevisiae and Preparation of DNA

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

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


INTRODUCTION

This protocol describes methods for isolation of DNA from a strain of S. cerevisiae carrying a recombinant YAC. Because the linear YAC DNAs are sensitive to shearing forces, pipettes with wide-bore tips should be used to transfer DNAs. The method is suitable for preparing DNA that will be used for agarose gel electrophoresis, Southern blotting, subcloning, genomic library construction, PCR, or other methods that do not require intact high-molecular-weight DNA. The expected yield from a 10-ml culture is 2-4 µg of yeast DNA.


MATERIALS

recipe caution Ammonium acetate (10 M)

Optional, please see Step 3.

Ethanol

Optional, please see Step 5.

caution Phenol:chloroform . . . [Full Text of this Article]


METHOD


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