With the support of our advanced protein engineering platform established for many years, Creative BioMart has developed a variety of random and degenerate mutagenesis methods. Our scientists provide you with a comprehensive customized random and degenerate mutagenesis library construction service.

Random mutagenesis is a technique that allows researchers to develop large libraries of specific DNA sequence variations. It is a prerequisite for the preparation of random DNA libraries, which is necessary for many genetic selections. These libraries can be used for protein structure, protein function and directed evolution research. Conventional in vivo random mutagenesis uses chemical mutagens (such as 2-aminopurine, 5-bromouracil, ethylmethane sulfonate) and the use of bacterial strains carrying single or multiple mutations in DNA to repair two methods. One of the most popular in vitro mutagenesis strategies is the error-prone polymerase chain reaction (PCR), which is based on errors in the incorporation of nucleotides into newly synthesized PCR products.

Random mutagenesis is different from other mutagenesis techniques in that it does not require researchers to have any prior knowledge of the structural properties of the target DNA sequence, and can unbiasedly discover new or beneficial mutations. Therefore, random mutation is particularly useful in the study of protein evolution.

Fig 1. Major steps in random mutagenesis by error-prone DNA polymerases. (Forloni M, et al., 2018 )

Methods of Random and Degenerate Mutagenesis

• The use of XL1-red: an engineered Escherichia coli with DNA repair deficiency, is one of the simplest mutagenesis and requires no subcloning step.
• Error-prone PCR: one of the most widely used random mutations. In the DNA amplification process, by changing the ratio of nucleotides and the concentration of divalent cations in the reaction, the mismatch of nucleotides can be promoted.
• DOP mutagenesis: It is a fast and effective method for random mutagenesis of small molecules of DNA and peptides. This method uses two chemically synthesized degenerate oligonucleotides as primers. By controlling the position and ratio of degraded nucleotides in the process of oligonucleotide synthesis, the position and mutation rate of the degraded area of the primer can be controlled.

Applications of Random and Degenerate Mutagenesis

• Protein expression and structure research.
• Enzyme kinetics or active site study.
• Antibody screening or antibody humanization research.

Service Principle

Creative BioMart is based on innovative, pragmatic and honest, adhering to the tenet of "Quality is our life, providing customers with the best quality service", providing customized services to customers around the world.

We will be glad to discuss details of intended interaction studies with you and develop experimental strategies/methods tailored to your requirement. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, Monday to Friday. Please do not hesitate to contact us for more information or to discuss in detail, if you are interested in our services.

References

1. Forloni M, Liu AY, et al.. (2018). Random Mutagenesis Using Error-Prone DNA Polymerases. Cold Spring Harb Protoc. 2018, 3.
2. Chusacultanachai S, Yuthavong Y. (2004) Random mutagenesis strategies for construction of large and diverse clone libraries of mutated DNA fragments. Methods Mol Biol. 270, 319-34.

• Homologous Recombination Library Construction Service
• Scanning Mutagenesis Library Construction Service
• Site-Directed Mutagenesis Library Construction Service