Protocol

Synthesis of Single-stranded DNA Probes of Heterogeneous Length from Bacteriophage M13 Templates

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

The following technique yields a heterogeneous population of short radiolabeled molecules 200-300 nucleotides in length. These probes are synthesized, as in Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates, by extension of an oligonucleotide primer on a single-stranded DNA template. The radiolabeled products of the reaction are then separated from the template by electrophoresis through a denaturing gel from which they are eluted directly into hybridization buffer. The method is useful for synthesizing single-stranded DNA probes of very high specific activity for Southern analysis of single-copy genes in complex genomes or for northern analysis of . . . [Full Text of this Article]

MATERIALS

METHOD