Site-Directed Mutagenesis is a powerful molecular biology technique used to introduce specific changes in the DNA sequence of a gene. It allows researchers to create precise mutations at desired locations in the genome, enabling the study of gene function, protein structure, and various biological processes.
By manipulating the DNA sequence, researchers can investigate the impact of specific mutations on protein function, study disease-causing mutations, and even engineer new proteins with improved properties. This technique has revolutionized the field of molecular biology and has become an indispensable tool for genetic engineering projects.
Overview of site-directed mutagenesis method using mix antibiotic selection(PE Carrigan, et al.,2011)
Services Offered
Products Offered
Service Process
FAQs
Services Offered
There are several types of Site-Directed Mutagenesis techniques, each with its own advantages and applications.
| Site-Directed Mutagenesis Type |
Applications |
Description |
| QuikChange Mutagenesis |
- Protein engineering, functional studies. |
- Description: Utilizes a unique DNA polymerase with 3' to 5' exonuclease activity to introduce site-specific mutations in a short and efficient PCR-based mutagenesis protocol. |
| Overlap Extension PCR (OE-PCR) |
- Protein structure-function studies, domain swapping. |
- Description: Involves the amplification of two DNA fragments with overlapping ends, followed by a second PCR to generate a full-length product containing the desired mutation. |
| Inverse PCR (iPCR) |
- Modification of circular DNA, plasmid engineering. |
- Description: Amplifies a circular DNA template with primers designed to include the desired mutation, followed by self-ligation to produce a modified circular DNA product. |
| SiteSat PCR |
- Amino acid scanning, investigating the effects of specific mutations. |
- Description: Systematically introduces mutations at individual amino acid positions within a target gene to analyze their impact on protein structure and function. |
| QuikChange Lightning |
- Fast and high-throughput mutagenesis. |
- Description: An optimized version of QuikChange Mutagenesis that reduces reaction times, allowing for quicker and more high-throughput generation of site-directed mutants. |
| Random Mutagenesis Kit |
- Random mutagenesis for protein evolution. |
- Description: Utilizes error-prone PCR to introduce random mutations across the entire target gene, providing a diverse pool of mutants for subsequent screening and selection. |
These Site-Directed Mutagenesis techniques find applications in various research areas. For example, in protein engineering, researchers can use this technique to introduce specific amino acid changes in a protein to improve its stability, activity, or binding affinity. In gene expression studies, Site-Directed Mutagenesis can be used to create reporter constructs or modify regulatory regions to study gene regulation. Additionally, in drug development, this technique can be employed to investigate the impact of specific mutations on drug efficacy or resistance.
If you have any further questions or require more information about our Site-Directed Mutagenesis service, please don't hesitate to contact us. We are committed to providing high-quality services and are eager to assist you in your genetic engineering endeavors.
Service Process
Our company offers comprehensive Site-Directed Mutagenesis services to assist researchers in their genetic engineering projects. We understand the importance of precise and reliable results, and our team of experts is dedicated to ensuring the success of your experiments. Our service process includes the following steps:
- Consultation: Our team of experts will discuss your project requirements and provide personalized advice on experimental design and strategy. We will work closely with you to understand your objectives and develop a tailored approach for your specific needs.
- Design and Primer Synthesis: Based on the target gene sequence and desired mutations, our experts will design the appropriate mutagenesis primers. These primers are crucial for introducing the desired changes in the gene sequence. We will then synthesize the primers using high-quality reagents and state-of-the-art technologies.
- Mutagenesis Reaction: Using our expertise in Site-Directed Mutagenesis techniques, we will perform the mutagenesis reaction to introduce the desired changes in the gene sequence. We utilize optimized protocols and advanced molecular biology tools to ensure efficient and accurate mutagenesis.
- Verification: After the mutagenesis reaction, we will verify the success of the introduced mutations. This includes DNA sequencing to confirm the presence of the desired changes and other analytical methods to assess the quality and integrity of the mutated gene construct. Our stringent quality control measures ensure reliable and accurate results.
- Delivery: Once the mutagenesis is confirmed, we will deliver the final mutated gene construct or plasmid to you. Along with the deliverables, we provide detailed documentation of the experimental procedures and results, enabling you to understand and replicate the experiments conducted.
Our goal is to provide you with a seamless and efficient experience, allowing you to focus on your research objectives while we handle the technical aspects of Site-Directed Mutagenesis.
FAQs
Q: How long does the Site-Directed Mutagenesis process typically take?
A: The timeline can vary depending on the complexity of the project and the specific requirements. However, our experienced team aims to deliver results within a reasonable timeframe. We understand the importance of timely results and will work closely with you to establish realistic timelines for your project.
Q: Can you assist with designing the mutagenesis primers?
A: Absolutely! Our experts have extensive knowledge in primer design and can provide guidance to ensure optimal results. By considering factors such as primer length, melting temperature, and specificity, we will design primers that are tailored to your specific target gene and desired mutations.
Q: What types of mutations can be introduced using Site-Directed Mutagenesis?
A: Site-Directed Mutagenesis allows for a wide range of mutations, including base substitutions, insertions, deletions, and more. Whether you need to introduce a single amino acid change or delete an entire region, our Site-Directed Mutagenesis services can accommodate your requirements.
Q: Is there a minimum order requirement for your Site-Directed Mutagenesis service?
A: We do not have a minimum order requirement. We understand that genetic engineering projects can vary in scale, from small-scale experiments to large-scale studies. Whether you require mutagenesis for a single gene or multiple genes, we are here to support your research needs.
Q: Can you work with non-standard organisms or difficult templates?
A: Yes, our team has extensive experience working with various organisms and challenging templates. We have successfully performed Site-Directed Mutagenesis on a wide range of organisms, including bacteria, yeast, plants, and mammalian cells. If you have non-standard organisms or difficult templates, please contact us to discuss your specific requirements. Our experts will develop customized solutions to meet your unique research needs.
Q: What are the advantages of Site-Directed Mutagenesis over other mutation techniques?
A: Site-Directed Mutagenesis offers several advantages, including the ability to introduce precise and specific mutations at desired locations in the genome. This technique allows for the study of gene function and protein structure with high accuracy. Additionally, Site-Directed Mutagenesis enables researchers to investigate the impact of specific mutations on biological processes and perform targeted genetic engineering.
Q: Can Site-Directed Mutagenesis be used for site-specific gene editing?
A: Yes, Site-Directed Mutagenesis can be used for site-specific gene editing. By introducing specific changes in the DNA sequence of a gene, researchers can modify or delete target gene regions with precision. This technique is especially useful for studying gene function and exploring the effects of specific mutations.
Q: Are there any limitations or challenges associated with Site-Directed Mutagenesis?
A: While Site-Directed Mutagenesis is a powerful technique, it does have some limitations. The success rate of mutagenesis reactions can vary depending on the complexity of the desired changes and the target gene sequence. Some gene regions may be more challenging to mutate than others. Additionally, the size of the gene and the presence of repetitive sequences can pose challenges during mutagenesis. However, with careful experimental design and optimization, many of these limitations can be overcome.
Q: Can Site-Directed Mutagenesis be used for large-scale mutagenesis projects?
A: Yes, Site-Directed Mutagenesis can be scaled up for large-scale projects. Whether you need to introduce mutations in a single gene or multiple genes, our Site-Directed Mutagenesis services can accommodate your requirements. Our team of experts can assist you in designing and executing mutagenesis experiments on a larger scale, ensuring accurate and reliable results.
Q: How can I request a quote for your Site-Directed Mutagenesis service?
A: To request a quote for our Site-Directed Mutagenesis service, please reach out to our team via email or phone. Provide us with details about your project, including the target gene, desired mutations, and any specific requirements. Our experts will promptly respond to your inquiry and provide you with a customized quote based on your needs.
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