Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot3225

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Isolation of High-molecular-weight DNA from Mammalian Cells Using Formamide

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 procedure, although lengthy, generates preparations of DNA that are large enough (200 kb) to be used for the construction of genomic libraries in high-capacity vectors and for analysis by pulsed-field electrophoresis. However, the concentration of DNA in the final preparation is low (approx. 10 µg/ml), as is the yield (approx. 1 mg of DNA/108 cultured aneuploid mammalian cells, such as HeLa cells).


MATERIALS

recipe Dialysis buffer I

recipe Dialysis buffer II

recipe Formamide denaturation buffer

Linear monomers and . . . [Full Text of this Article]


METHOD


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[Abstract] [Full Text] [PDF]