Long DNA Synthesis

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Overview
While traditional chemical methods limit DNA lengths to 200 nucleotides, the demand for kilobase to megabase-level genes, even entire genomes, necessitates the support of advanced in vitro assembly technologies. Tsingke stands at the forefront of overcoming key challenges in genome synthesis, particularly focusing on advancing long DNA synthesis and 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. We've also achieved industrial-scale production of 50 kb large DNA fragment, with demonstrated capabilities to deliver 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, while also supporting DNA digital storage and fundamental life science research. Through our technological advancements, we aim to tackle pressing issues such as food scarcity, drug development, disease treatment optimization, environmental challenges, and innovative 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
What is delivered for long DNA synthesis services?
We provide one tube of lyophilized plasmid (stored at -20°C).
On which vectors are the synthesized large fragments cloned? Can specific vectors be used?
Our company defines long DNA sequences as those exceeding 10 kb. For sequences ranging from 10 kb to 30 kb, yeast assembly methods are used to first construct them into the pcc1413 vector before transferring them to your specified target vector. Sequences exceeding 30 kb will be directly constructed into the pcc1413 vector.
**For Research Use Only. Not for use in diagnostic procedures.
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