Protocol

Random Priming: Radiolabeling of Purified DNA Fragments by Extension of Random Oligonucleotides

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

Short oligonucleotides of random sequence can serve as primers for the initiation of DNA synthesis at multiple sites on single-stranded DNA templates. During extension of the primers by DNA polymerase, the complement of every nucleotide in the template (except those at the extreme 5' terminus) will be incorporated into the product at approximately equal frequency. The DNA synthesized can be labeled by using one [-³²P]dNTP . . . [Full Text of this Article]

MATERIALS

METHOD