Laboratory Methods in Enzymology: DNA

Edited by

  • Jon Lorsch, Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA

Methods in Enzymology volumes provide an indispensable tool for the researcher. Each volume is carefully written and edited by experts to contain state-of-the-art reviews and step-by-step protocols.

In this volume, we have brought together a number of core protocols concentrating on DNA, complementing the traditional content that is found in past, present and future Methods in Enzymology volumes.

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Audience

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

 

Book information

  • Published: September 2013
  • Imprint: ACADEMIC PRESS
  • ISBN: 978-0-12-418687-3


Table of Contents

Explanatory chapter: PCR Primer design

Explanatory Chapter: How Plasmid Preparation Kits Work

Explanatory Chapter: Introducing Exogenous DNA into cells

Agarose Gel Electrophoresis

Analysis of DNA by Southern Blotting

Purification of DNA Oligos by Denaturing Polyacrylamide Gel Electrophoresis (PAGE)

Molecular Cloning

Rapid creation of stable mammalian cell lines for regulated expression of proteins using the Gateway® Recombination Cloning Technology and Flp-In T-REx® lines

Restrictionless cloning

Isolation of plasmid DNA from bacteria

Preparation of Genomic DNA from Bacteria

Preparation of Genomic DNA from Saccharomyces cerevisiae

Isolation of Genomic DNA from Mammalian Cells

Sanger Dideoxy Sequencing of DNA

Preparation of fragment libraries for Next-Generation Sequencing on the Applied Biosystems SOLiD platform

Explanatory Chapter: Next Generation Sequencing

Generating mammalian stable cell lines by electroporation

Transient mammalian cell Transfection with Polyethylenimine (PEI)

Site-Directed Mutagenesis

PCR-based random mutagenesis

Megaprimer Method for Mutagenesis of DNA

Explanatory Chapter: Troubleshooting PCR

Explanatory Chapter: Quantitative PCR

General PCR

Colony PCR

Chemical Transformation of Yeast

Transformation of E. coli via electroporation

Transformation of Chemically Competent E. coli