ProLongGene

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Overview
Tsingke stands at the forefront of overcoming key challenges in genetic synthesis, with a focus on the development of long fragment synthesis and assembly technologies. In the realm of gene synthesis, where chemical methods limit lengths to 200 nucleotides, achieving kilobase to megabase-level genes, even entire genomes, necessitates the support of advanced in vitro assembly technologies.
 
With years of expertise and ongoing innovation, we've transcended the boundaries of genetic synthesis. Our breakthroughs include mastering the core technology of yeast assembly for multiple segments and large fragments, enabling precise and efficient one-step assembly of DNA fragments in Saccharomyces cerevisiae. Our proficiency extends to the industrial-scale production of 50 kb large fragment DNA, with a demonstrated capability of delivering fragments as long as 200 kb.
 
Our Gene Factory boasts state-of-the-art long fragment synthesis and assembly technologies. We are dedicated to providing customized solutions for metabolic pathway synthesis, genome synthesis, and assembly, supporting DNA digital storage and fundamental life science research. Our technological innovations offer solutions to address food scarcity, develop novel drugs, optimize disease treatment methods, resolve environmental issues, and explore more ideal data storage solutions.
Advantages
Up to 200kb
Achieved construction as long as 200 kb
High Assembly Efficiency
One-step assembly of DNA fragments in Saccharomyces Cerevisiae assembly system
Accuracy
Guaranteed with Sanger sequencing and NGS

Service Details

1

Length

Turnaround time

(Business Day)

Vector Deliverables
>10 kb Evaluation pCC1413

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

Sequencing map (.abl file);

Target sequence (.seq file);

COA Report(electronic). 

 

Workflow
sequence analysis & codon optimization
Sequence Optimization
sequence analysis & codon optimization
design & synthesis
Oligo Synthesis
design & synthesis
In Vitro assembly & In Vivo assembly
Construction
In Vitro assembly & In Vivo assembly
QC for NGS & Sanger sequencing
Quality Control
QC for NGS & Sanger sequencing
ship the plasmid containing your long DNA sequence insert
Delivery
ship the plasmid containing your long DNA sequence insert
Case
Figure 1. Long Fragment and Genome Synthesis Based on Saccharomyces Cerevisiae System

Currently, Tsingke has successfully developed the Spore Bacillus One-Step Assembly technology to replace traditional extracellular enzyme ligation methods. This technology allows for the rapid and efficient acquisition of 5 kb DNA sequences, enabling the simultaneous assembly of up to 10 fragments in a single step. This significantly shortens the synthesis cycle, reducing the time required by at least half compared to traditional methods. Additionally, we have optimized the yeast assembly system, greatly enhancing the success rate and transformation efficiency of multi-segment assembly. This optimization enables us to successfully complete the assembly of 200 kb DNA.
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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