Single-stranded DNA (ssDNA)

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Recent research highlights the effectiveness of single-stranded DNA (ssDNA) as a homology-directed repair donor template in CRISPR gene editing. Compared to double-stranded DNA (dsDNA), ssDNA significantly boosts editing efficiency, reduces off-target effects, and is versatile in various biological reactions, especially in DNA nanotechnology.
In CRISPR and CRISPR-Cas9 genome editing experiments, long ssDNA serves as a potent donor template, enhancing both insertion and gene replacement efficiency. Its applications extend to single-strand conformation polymorphism, in vitro transcription studies, nucleic acid enzyme S1 mapping, probe preparation, labeling, and differential hybridization.
Beyond experiments, ssDNA plays a role in DNA nanotechnology, serving as a scaffold for drug delivery, molecular diagnostics, DNA-based data storage, and diverse nanoscale applications. With a lower risk of random integration, ssDNA is particularly suitable for gene editing in primary cells, stem cells, and the creation of genetically modified animal models.
Tsingke now provides top-notch, sequence-validated ssDNA to enhance the efficiency of your CRISPR experiments.
Wide Synthesis Range
Offering the synthesis of ssDNA
up to 6000 nt
Low Cytotoxicity
Achieve higher homologous recombination and knock-in efficiency, minimizing cytotoxic effects

High Editing Efficiency
9-fold higher CRISPR/Cas9 gene knock-in efficiency than dsDNA as a homologous recombination template

Service Details

Length (nt)

Turnaround time (Business Day)




Lyophilized Product Powder; 
Sequencing map (.abl file);
Target sequence (.seq file);
COA Report(electronic).











sequence analysis & codon optimization
Sequence Optimization
sequence analysis & codon optimization
design & synthesis
Oligo Synthesis
design & synthesis
ssDNA synthesis
Single-Stranded DNA Synthesis
ssDNA synthesis
Sanger sequencing
Quality Control
Sanger sequencing
ship the lyophilized powder
ship the lyophilized powder
Figure 1: 94% Purity Test
Figure 2: 99.3% Purity Test
Related Resource
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 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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