Protocol

Random Priming: Radiolabeling of DNA by Extension of Random Oligonucleotides in the Presence of Melted Agarose

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

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INTRODUCTION

A variation of the technique described in Random Priming: Radiolabeling of Purified DNA Fragments by Extension of Random Oligonucleotidescan be used to radiolabel DNA in slices cut from gels cast with low-melting-temperature agarose. For details on separating labeled probe from unincorporated dNTPs, please see Appendix 8 in the print version of the manual.

MATERIALS

5x Oligonucleotide labeling buffer

Ammonium acetate (10 M)

Optional, please see Step 5.

Bovine serum albumin . . . [Full Text of this Article]

METHOD