Mutagenesis

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Overview
Site-directed mutagenesis, a molecular biology technique, could deliberately introduce precise changes or mutations into a specific DNA sequence. This includes base insertions, deletions, and point mutations. Now Site-directed mutagenesis serves as a powerful tool to efficiently enhance the expression, traits, and characterization of the targeted protein. The technology of site-directed mutagenesis is now extensively employed in the investigation of protein functional site structures, optimization of enzyme activity, understanding of DNA component functions and interactions, as well as applications in gene therapy and other research areas.

Tsingke provides specialized and cost-effective site-directed mutagenesis services, precisely introducing your requirement mutations for any gene template and any target number of mutations. This facilitates the synthesis of increased experimental efficiency and cost-effectiveness for your research endeavors.
Advantages
Accuracy
Guaranteed with Sanger sequencing and NGS
Unlimited Sites
Introduce your mutations at any site
Cost-effective
All mutations found within a 30-base region will be defined as one whole mutation
Service Details

Length

*Turnaround time

(Business Day)

Deliverables

<3 kb

7~10

1 tube of lyophilized plasmid DNA
 (about 1~4 μg/ tube); 

Sequencing map (.abl file);

Target sequence (.seq file);

COA Report(electronic). 

3 kb~5 kb

10~12

5 kb~7 kb

12~15

>7 kb

Evaluation

*Note: Get your accurate estimated turnaround time by emailing gene@tsingke.com.cn
Workflow
oligo design & synthesis
Order Design
oligo design & synthesis
PCR amplification or digestion
Sequence Acquisition
PCR amplification or digestion
linearized the purified DNA fragment with the cut destination vector
Vector Construction
linearized the purified DNA fragment with the cut destination vector
QC for NGS & Sanger sequencing
Quality Control
QC for NGS & Sanger sequencing
ship the plasmid containing your target gene fragment
Delivery
ship the plasmid containing your target gene fragment
Related Resource
FAQ
Does Tsingke provide codon optimization? Does optimization have an impact on gene expression?
Tsingke offers free codon optimization.
For the same amino acid in different hosts, the corresponding codons may exhibit varying degrees of preference. Codon optimization involves utilizing preferred codons and avoiding rare codons to optimize the expression of a sequence in a specified host. There is a strong correlation between gene expression levels and codon preference. Tsingke's unique GeneOptimizer software utilizes a codon optimization algorithm that supports optimization for tens of thousands of hosts and provides customized services. The main parameters for optimization include codon preference, GC content, restriction enzyme sites (deletion), poly structure, sequence repeats, etc.
Which termination codon is preferred in the E. coli expression system? What about in mammalian systems?
In E. coli, TAG is rarely used; TAA is frequently used; TGA is also available.
In mammals, TGA is used more frequently than TAA and TAG.
Relative to the differences between mammals and E. coli, codon usage tables do not differ very much between different mammalian organisms.
How does Tsingke solve high-difficulty genes, such as those with high or low GC content, highly repetitive sequences, very long sequences, and special
High and low GC content, repetitive sequence:
Solution: Divide the sequence into small segments to reduce the impact of structure on synthesis, and choose the optimal connection solution, such as enzyme digestion and ligation and multi-segment recombination.
Very long sequences:
Solution: use multi-segment recombination to construct divided small segments, and combine them with vitro and vivo assembly.
Special parts of vectors (e.g. ccdB, LTR):
Solution: Use different competent cells corresponding to different parts of vectors, such as DB3.1, which renders the strain resistant to the toxic effects of the ccdB gene, the recombination-deficient Stbl series, etc.
Other unpredictable difficulties, such as genetic instability and toxicity, and the introduction of random mutations and deletions
Solution: Try different competent cells.
Can you keep my sequence information confidential?
Yes, all customer information will be kept confidential.
Which sites in the pUC57 vector will the synthesized gene be cloned?
Generally, the default clone is between EcoRⅠ and Hind Ⅲ. If you have particular needs, please note and we can select other suitable cloning sites according to the experimental needs based on your sequence.
Are there any requirements for my provided-vector?
We need you to ship at least 2 μg plasmid.
What vector does Tsingke use by default? Can I send my vectors to you?What vector does Tsingke use by default? Can I send my vectors to you?
We provide pUC57 vectors containing Amp (ampicillin) for free by default, and we also have a vector library for 160+ options that you could use for free.
Alternatively, you may use your own vector. Simply provide the vector sequence and a vector sample. It is recommended to ship your vector in advance, otherwise, the Turnaround Time (TAT)  may need to be extended by 3 business days to verify the vector.
What information do you need to prepare a quote?
We require information on the insert DNA sequence or amino acid sequence, and the host species if codon optimization is needed. Please ensure that the restriction sites are specified at the 5'/3' end or indicate if any internal modifications need to be avoided. Additionally, let us know if you wish to include other elements in the sequence, such as the Kozak sequence, stop codon, etc. 
Feel free to complete the evaluation form and send it via email to info@tsingke.com.cn. For any inquiries or project requirements, you can contact our account managers or sales managers through email. Our technical support team will promptly assess the price and turnaround time (TAT) and provide you with a detailed quotation as soon as possible."
**For Research Use Only. Not for use in diagnostic procedures.
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