Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4035

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Retrieval of DNA Fragments from Pulsed-field Gels following DNA Concentration

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

DNA contained in a slice of low-melting-temperature agarose is first concentrated by electrophoresis into a high-percentage agarose gel, and then isolated by treatment with agarase. The resulting DNA preparation is purified by microdialysis.


MATERIALS

Agarase

recipe Agarase digestion buffer

Use the buffer supplied with the enzyme. It is critical that all buffers be supplemented with 100 mM NaCl, 30 µM spermine, and 70 µM spermidine to enhance recovery of the DNA.

DNA size standards

Please see Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of Intact DNA from Yeastand Markers for Pulsed-field Gel Electrophoresisfor the preparation of two different ranges . . . [Full Text of this Article]


METHOD


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