Protocol

Mixed Oligonucleotide-primed Amplification of cDNA (MOPAC)

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

In MOPAC, the amino-terminal and carboxy-terminal sequences of a peptide are used to design two redundant families of oligonucleotides encoding the amino- and carboxy-terminal sequences of the peptide. The primers are used either to amplify a segment of cDNA prepared by RT-PCR from a tissue known to express the protein or to amplify a segment of DNA from an established genomic or cDNA library. Because the length of the peptide is known, the size of the expected PCR product can be predicted exactly. After gel electrophoresis to resolve the amplification products, DNAs of the correct size are isolated, cloned, and . . . [Full Text of this Article]

MATERIALS

METHOD