Chapter title |
An In Vitro Single-Primer Site-Directed Mutagenesis Method for Use in Biotechnology.
|
---|---|
Chapter number | 26 |
Book title |
In Vitro Mutagenesis
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6472-7_26 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6470-3, 978-1-4939-6472-7
|
Authors |
Yanchao Huang, Likui Zhang |
Editors |
Andrew Reeves |
Abstract |
Site-directed mutagenesis is a powerful method to introduce mutation(s) into DNA sequences. A number of methods have been developed over the years with a main goal being to create a high number of mutant genes. The single-mutagenic primer method for site-directed mutagenesis is the most direct method that yields mutant genes in about 25-50 % of transformants in a robust, low-cost reaction. The supercompetent XL10-Gold bacteria used in the Stratagene protocol carry a phage, which may be a problem for some applications; however, in our single-mutagenic primer method the supercompetent bacteria are not needed. A thermostable DNA polymerase with high fidelity and processivity, such as Phusion DNA polymerase, is required for our optimized procedure to avoid extra mutation(s) and enhance mutagenic efficiency. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 14 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 5 | 36% |
Researcher | 2 | 14% |
Student > Doctoral Student | 1 | 7% |
Student > Master | 1 | 7% |
Unknown | 5 | 36% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 5 | 36% |
Environmental Science | 1 | 7% |
Agricultural and Biological Sciences | 1 | 7% |
Immunology and Microbiology | 1 | 7% |
Unknown | 6 | 43% |