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A rapid, simple, and effective method of constructing a randomly mutagenized plasmid library free from ligation
- Source :
- Bioscience, biotechnology, and biochemistry. 72(4)
- Publication Year :
- 2008
-
Abstract
- The QuikChange site-directed mutagenesis methodology was applied to constructing a randomly mutagenized plasmid library simply by adding manganese to the reaction mixture. This method is superior to the normally employed Pol I-type polymerase-based error-prone PCR in that (i) it does not require a subsequent ligation reaction, and (ii) there is no accumulation of mutations at the same site. alpha-Complementation analysis and subsequent sequence analyses of the lacZ alpha genes in the mutated library revealed that the mutations occurred randomly within the target gene and involved all possible base substitutions.
- Subjects :
- Time Factors
Mutagenesis (molecular biology technique)
Biology
medicine.disease_cause
Applied Microbiology and Biotechnology
Biochemistry
Analytical Chemistry
Plasmid
medicine
Genomic library
Molecular Biology
Gene
Polymerase
Gene Library
Genetics
Mutation
Manganese
Stochastic Processes
Base Sequence
Organic Chemistry
General Medicine
Nucleotidyltransferase
Molecular biology
biology.protein
Mutagenesis, Site-Directed
Ligation
Biotechnology
Plasmids
Subjects
Details
- ISSN :
- 13476947
- Volume :
- 72
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- Bioscience, biotechnology, and biochemistry
- Accession number :
- edsair.doi.dedup.....aee45a841863ff1fe5d8bb015f3b88e1