Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3842

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protocolProtocol

Quantitative 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

Quantitative PCR involves co-amplification of two templates: a constant amount of a preparation containing the desired target sequence and serial dilutions of a reference template that is added in known amounts to a series of amplification reactions. The concentration of the target sequence is determined by simple interpolation into a standard curve. To reduce the chance of contamination with exogenous DNAs, prepare and use a special set of reagents and solutions for PCR only. Bake all glassware for 6 hours at 150°C and autoclave all plasticware.


MATERIALS

recipe 10x Amplification buffer

Include 0.01% (w/v) gelatin in the buffer.

Antisense primer (20 µM) . . . [Full Text of this Article]


METHOD


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