Our promise to you:
Guaranteed product quality, expert customer support.
24x7 CUSTOMER SERVICE
CONTACT US TO ORDER
Gene mutagenesis is a key technology in molecular biology and genetic engineering, enabling precise, controllable changes at specific DNA sites. It is essential for studying gene function, protein engineering, functional genomics, disease models, and industrial enzyme optimization. Creative Biogene leverages advanced platforms, mature strategies, and high-fidelity systems to provide efficient, accurate, and flexible one-stop gene mutagenesis solutions.
Our services cover single-site and multi-site mutations, deletions, insertions, and random mutagenesis libraries. From small functional validation projects to large-scale high-throughput screening and directed evolution campaigns, we deliver highly accurate, customized constructs quickly, ensuring clients receive the materials they need without delay.
Figure 1. Schematic of Mutant Construction
Site-Directed Mutagenesis
Based on a high-fidelity PCR mutagenesis platform, we can achieve single-base substitutions, multi-site combinatorial mutations, sequence deletions, short fragment insertions, and domain replacements. This approach does not require full de novo DNA synthesis. Instead, using either a client-provided or our synthesized template, pre-designed mutation primers introduce targeted changes into the sequence, which are then cloned into the desired vector. Final constructs undergo 100% sequence verification to ensure accuracy.
Random Mutagenesis
We employ Error-Prone PCR to deliberately reduce DNA replication fidelity during amplification by adjusting reaction conditions (Mg²⁺, Mn²⁺, dNTP ratios, polymerase type and concentration, pH, etc.), generating random base mismatches in the target gene. Our optimized system can introduce 1–4 random amino acid substitutions per 1000 bp, covering the entire ORF or selected fragments.
Creative Biogene's gene mutagenesis services follow a highly standardized workflow, providing full project management from initial assessment to final product delivery:
1Project Intake and Assessment: Upon receiving target gene information from the client (sequence, mutation sites or regions, vector details), we evaluate sequence feasibility and provide an optimized design plan.
2Primer and Strategy Design: Using professional software, primers are designed to ensure high specificity and avoid dimers or secondary structure interference, with special optimization for long fragments or high GC regions.
3Mutagenesis Reaction and Construction: PCR reactions are performed using high-fidelity or low-fidelity (for random mutagenesis) polymerase systems to introduce the desired mutations. The resulting products are cloned into the client-specified or provided vector.
4Sequence Verification and Quality Control: All constructs undergo bidirectional sequencing to ensure no off-target mutations and complete agreement with the design.
5Delivery and Documentation: We provide lyophilized plasmid DNA, recombinant plasmid-containing stab cultures, or glycerol stocks, accompanied by a complete COA, sequencing chromatograms, and sequence files.
Creative Biogene's gene mutagenesis services uphold the highest quality control and delivery standards, ensuring 100% mutation accuracy through the use of high-fidelity enzymes and bidirectional sequencing, with all constructs verified by colony PCR and plasmid extraction to prevent vector background contamination. Every step of the process is fully traceable, with experimental logs, reagent batches, and sequencing data archived, and we offer flexible delivery options, including glycerol stocks, stab cultures, or purified plasmids, accommodating special buffer storage requirements.
Creative Biogene's services provide clear advantages for a wide range of research needs, supporting comprehensive mutation types such as single-site, multi-site, deletions, insertions, and random libraries, with precise mutagenesis achievable for DNA fragments up to 12 kb. As a one-stop, end-to-end solution, we also offer optional gene synthesis, vector construction, and expression testing, delivering standard-length sequences within 5–8 working days, while our optimized systems reduce reagent costs, ensuring the highest quality and cost-effective results for every project.
For inquiries about our gene mutagenesis services, project consultation, or custom solutions, please reach out to our scientific support team. We are ready to provide detailed guidance, help design your mutagenesis strategy, and ensure the fastest and most accurate delivery of your research materials.
Q1: What are your mutation efficiency and accuracy?
A1: Site-directed mutagenesis has a nearly 100% success rate, with all constructs confirmed by bidirectional sequencing. Random mutagenesis library diversity and mutation rates can be customized according to client needs (1–4 amino acid substitutions per 1000 bp is typical).
Q2: Can you perform mutagenesis on high-GC or long DNA fragments?
A2: Yes. We use special primer design and additive systems for high-GC sequences, supporting precise mutagenesis of DNA sequences up to 12 kb.
Q3: Will Error-Prone PCR introduce too many irrelevant mutations?
A3: Mutation frequency and bias are adjusted according to client requirements, and library quality can be analyzed by next-generation sequencing (NGS) to balance diversity and functional integrity.
Q4: How should delivered materials be stored and handled?
A4: Lyophilized plasmids can be transported at room temperature and stored at –20°C upon receipt. Glycerol stocks require –80°C for long-term storage, while stab cultures can be stored at 4°C for short-term and revived promptly.
Q5:What PCR-based site-directed mutagenesis methods are available?
A5: Methods:
1. Overlap Extension PCR – Two-step PCR to introduce mutations. Labor-intensive, sensitive to conditions, risk of template contamination.
2. Rolling Circle Amplification – Amplifies plasmid with mutation primers, ligation, and template digestion. Challenging for long plasmids; multi-site mutations require more time and reagents.
3. Homologous Recombination – Uses homologous arms in primers for recombination after template removal. Supports single/multi-site mutations, insertions, deletions, and long fragments with high efficiency and lower effort.
Comparison:
| Method | Difficulty | Time | Gene Length | Efficiency | Scope |
| Overlap Extension PCR | ★★★★★ | ★★★★★ | ★★ | ★★ | Single/Multi-site |
| Rolling Circle | ★★★ | ★★★ | ★★★ | ★★★ | Single/Multi-site |
| Homologous Recombination | ★★★ | ★★★ | ★★★★★ | ★★★★★ | Single/Multi-site/Insertion/Deletion |
Copyright © Creative Biogene. All rights reserved.