Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot3865

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Rapid Amplification of 3' cDNA Ends (3'-RACE)

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

3'-RACE reactions are used to isolate unknown 3' sequences or to map the 3' termini of mRNAs onto a gene sequence. 3'-RACE requires knowledge of a small region of sequence within either the target RNA or a partial clone of cDNA. A population of mRNAs is transcribed into cDNA with an adaptor-primer consisting at its 3' end of a poly(T) tract and at its 5' end of an arbitrary sequence of 30-40 nucleotides. Reverse transcription is usually followed by two successive PCRs. The first is primed by a gene-specific sense oligonucleotide and an antisense primer complementary to the arbitrary sequence . . . [Full Text of this Article]


MATERIALS


METHOD


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