Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3844

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Differential Display-PCR

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

This method uses PCR to amplify and display many cDNAs derived from the mRNAs of a given cell or tissue type. The method relies on two different types of synthetic oligonucleotides: anchored antisense primers and arbitrary sense primers. A typical anchored primer is complementary to approx. 13 nucleotides of the poly(A) tail of mRNA and the adjacent two nucleotides of the transcribed sequence. Anchored primers therefore anneal to the junction between the poly(A) tail and the 3'-untranslated region of mRNA templates, from where they can prime synthesis of first-strand cDNA. A second primer, an arbitrary sequence of approx. 10 nucleotides, . . . [Full Text of this Article]


MATERIALS


METHOD


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