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In-vitro Mutagenesis

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: Apr 26, 2026

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

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

Published on: May 15, 2018

[A method for introducing mutations into large vectors].

Fanrong Meng¹, Chen Chen¹, Haisu Wan¹

¹Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenviroment, Tianjin Lung Cancer Institute,  Tianjin Medical University General Hospital, Tianjin 300052, China.

Zhongguo Fei AI Za Zhi = Chinese Journal of Lung Cancer

|July 19, 2014

Summary

This summary is machine-generated.

This study presents a new site-directed mutagenesis method for efficiently introducing mutations into large vectors (>9 kb) with over 90% success rates, offering a valuable tool for molecular biology research.

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Last Updated: Apr 26, 2026

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

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Published on: May 15, 2018

A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors

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Published on: October 30, 2016

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

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

Published on: February 5, 2019

Area of Science:

Molecular Biology
Genetic Engineering

Background:

Site-directed mutagenesis is crucial in molecular biology.
Existing methods are often unsuitable for large vectors.

Purpose of the Study:

To develop an effective method for site-directed mutagenesis in large vectors.
To overcome limitations of current techniques for large DNA constructs.

Main Methods:

A novel protocol involving vector primers, bridge primers, and type IIs endonuclease digestion.
PCR amplification followed by fragment ligation to introduce specific mutations.
Utilizes complementary oligonucleotides to form a mutagenic fragment.

Main Results:

Successfully introduced mutations into vectors exceeding 9 kb in size.
Achieved mutation rates greater than 90% with the developed protocol.
Demonstrated high efficiency for targeted genetic modifications.

Conclusions:

The described method is a powerful tool for site-directed mutagenesis in large vectors.
Provides a reliable and efficient approach for genetic manipulation of large DNA constructs.
Enhances capabilities for molecular biology research involving large vectors.