Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4037

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Isolation of DNA from Mammalian Cells by Spooling

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 100 words of the full text of this article appear below.


INTRODUCTION

This procedure is used to prepare DNA simultaneously from many different types of samples or tissues. Although the DNA is generally too small (approx. 80 kb) for efficient construction of genomic DNA libraries, it gives excellent results in Southern hybridizations and PCRs. Cultured aneuploid mammalian cells (2 x 107, e.g., HeLa cells) yield 100 µg of DNA in a volume of 1 ml.


MATERIALS

Ethanol (room temperature)

Linear monomers and concatemers of bacteriophage {lambda} DNA (please see Markers for Pulsed-field Gel Electrophoresis)

Prepare {lambda} DNA as described in Extraction of Bacteriophage {lambda} DNA from Large-scale Cultures Using Proteinase K and SDSor . . . [Full Text of this Article]


METHOD


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