Mutagenesis Custom Order

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In DNA cloning, mutagenesis allows ones to introduce specific or random changes into a cloned DNA fragment, creating a variety of the gene for further study.

1. Site-directed Mutagenesis:

This method targets a specific location within the DNA sequence for a precise alteration. It's ideal when you want to introduce a well-defined change, like replacing a single amino acid in the encoded protein.

  • PCR with mutagenic primers: Primers used for PCR amplification can be specifically designed to incorporate the desired mutation at a specific location during DNA synthesis.
  • Restriction enzyme digestion and ligation: The cloned DNA fragment can be cut with restriction enzymes to create a linear piece. Synthetic oligonucleotides containing the desired mutation are then inserted using ligase enzymes to replace the original sequence.
  • Cas9-based methods: This approach utilizes the CRISPR-Cas9 system for precise genome editing. A guide RNA directs Cas9 nuclease to a specific location in the DNA, and a repair template containing the desired mutation is introduced for cellular repair machinery to incorporate the change.

2. Random Mutagenesis:

This method performs random mutagenesis throughout the cloned DNA fragment. It is useful for generating libraries of variants with different changes to allow selection of clones with desired characteristics.

  • Physical or chemical treatment: DNA is exposed to chemical or physical treatment to induce random mutations. This treatment may use oxidizing agents, alkylating agents, or methods such as UV and X-rays.
  • PCR-based mutagenesis: Mutations are induced by amplifying DNA using special PCR conditions that allow specific mutations to occur. By using a primer containing mixed bases, it is possible to induce random mutation only in specific regions.

Bioneer provides various types of mutagenesis services. Base substitution, deletion, or insertion of a specific region is possible. Additionally, mutagenesis services are available that allow tagging a reporter such as GFP to a specific gene and creating a fusion protein by linking multiple genes.

It's possible. Cloning service is available after mutagenesis, but the vector to be used for cloning must be provided separately.

  
  
  
  
  

Overview

Mutagenesis service provides mutant gene synthesis essential for researches based on not only protein structure and functions, but also improvements on enzymatic functions easily through Bioneer’s gene synthesis service.

Features and Benefits

  • Efficiency

    Save time and money through manufacturing by integrated production system from genetic raw material to high-speed oligo synthesizer

  • Accuracy

    100% guaranteed exact sequences using Automatic DNA sequencer (ABI 3730)

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