PCR Approaches to DNA Mutagenesis and Recombination: An Overview
Current polymerase chain reaction (PCR) innovations provide powerful tools for the cloning of previously unknown genes as well as characterization of their functions. Examples of the latter used include PCR-mediated in vitro mutagenesis and recombination of the cloned genes. PCR approaches have become the method of choice to generate arrays of predefined mutations or recombination within the gene of interest. In the postgenomic era, these mutants or recombinants are highly desirable for the study of functional genomics, gene expression, protein structure-function relationships, protein-protein interactions, protein engineering, and in vitro evolution of enzymes.
- DNA Methyltransferase Assays
- Analysis of Mitosis in Squash Preparations of Larval Brains: Orcein, Giemsa, Hoechst 33258, DAPI, Quinacrine, and N-Banding
- Analysis of Protein-DNA Binding by Streptavidin-Agarose Pulldown
- Chromosome Orientation Fluorescence In Situ Hybridization or Strand-Specific FISH
- l,lO-Phenanthroline-Copper Ion Nuclease Footprinting of DNA-Protein Complexes in Situ Following Mobility-Shift Electrophoresis A
- Forward Genetic Screens in Xenopus Using Transposon-Mediated Insertional Mutagenesis
- Detection of Reporter Gene Expression in Murine Airways
- Assays for the Preferential Binding of Human Topoisomerase I to Supercoiled DNA
- Use of Aptamer Tagging to Identify In Vivo Protein Binding Partners of Small Regulatory RNAs
- Constructing and Screening a cDNA Library: Methods for Identification and Characterization of Novel Genes Expressed Under Condit