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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
In vitro Mutagenesis01:16

In vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Related Experiment Video

Updated: May 9, 2026

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli
07:04

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Published on: February 5, 2019

PCR-mediated site-directed mutagenesis.

Michael F Carey, Craig L Peterson, Stephen T Smale

    Cold Spring Harbor Protocols
    |August 3, 2013
    PubMed
    Summary
    This summary is machine-generated.

    This study introduces a simplified method for creating point mutations in DNA using a single PCR step. This technique efficiently generates single or double mutations in proteins and promoter regions.

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    Last Updated: May 9, 2026

    Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli
    07:04

    Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

    Published on: February 5, 2019

    Homemade Site Directed Mutagenesis of Whole Plasmids
    07:11

    Homemade Site Directed Mutagenesis of Whole Plasmids

    Published on: May 11, 2009

    Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast
    07:18

    Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

    Published on: May 15, 2018

    Area of Science:

    • Molecular Biology
    • Protein Engineering
    • Genetic Engineering

    Background:

    • Traditional site-directed mutagenesis methods can be complex and time-consuming.
    • Efficient generation of specific DNA mutations is crucial for protein engineering and functional studies.

    Purpose of the Study:

    • To present a simplified protocol for generating point mutants.
    • To enable easy introduction of single or double mutations into proteins and promoter sites.

    Main Methods:

    • Utilizes a single polymerase chain reaction (PCR) step.
    • Employs full plasmid amplification for mutant generation.
    • Applies standard protein expression vectors and commercially available reagents.

    Main Results:

    • Successfully generates point mutants with a single PCR step.
    • Demonstrates suitability for introducing small mutations into promoter sites.
    • Shows high efficacy for introducing single or double mutations into proteins.

    Conclusions:

    • The described method offers an elegant and simple approach to mutagenesis.
    • This protocol is easily adaptable for use in standard molecular biology laboratories.
    • Facilitates efficient protein engineering and genetic modification studies.