Single-stranded DNA (ssDNA) is a nucleic acid composed of a single strand of deoxyribonucleotides. In nature, single-stranded DNA is found in certain viruses, such as the genome of adeno-associated viruses (AAVs), or as an intermediate in DNA replication and repair. Recently, its therapeutic potential has gained recognition.

Recently, single-stranded DNA is gaining prominence as a superior alternative to plasmid DNA (pDNA) or linear double-stranded DNA (dsDNA) for gene editing, vaccines, advanced therapies and diagnostics, and more.

Unlike double-stranded DNA, single-stranded DNA does not have complementary base pairing. Therefore, single-stranded DNA offers greater structural flexibility and can adopt various topologies and secondary structures. Additionally, no known cytosolic sensors specifically detect single-stranded DNA, allowing for safer and more repeatable treatments due to a lack of immunogenicity.

Gxstrands uses a fermentation-based manufacturing process with proven E. coli systems to efficiently produce flexible single-stranded DNA sequences with customizable topology (linear or circular) and lengths (up to 10kb) on a large scale, achieving gram-level quantities.

Previously, single-stranded DNA production was carried out in vitro by chemical synthesis or by enzymatic processes like rolling circle amplification.

GXstrands produces gram-level quantities and beyond. Our initial offering for testing is a 3L run, typically yielding 500 µg to 1 mg, depending on the specific sequence and its complexity. All material generated from the 3L run is shipped to the customer. Once the optimal sequence is identified, our fermentation-based approach enables scalable production to gram- or even kilogram-level quantities of single-stranded DNA.

Gxstrands single-stranded DNA sequences are available in virtually any length, up to 10 kb and beyond for research use only (RUO). GMP-like and GMP grades are in process. Contact info@gxstrands.com for more information.

We offer customizable production of single-stranded DNA in either sense or antisense orientation (only 1 species in final product). Tailored to your needs, we design the production plasmid, ensuring that the final single-stranded DNA product contains your preferred orientation.

The target single-stranded DNA sequence is verified through Sanger sequencing. Purity is assessed using gel electrophoresis. Endotoxins are removed to a concentration of less than 0.5 EU/mL. Additional tests can be incorporated for specific projects upon request. Contact info@gxstrands.com to discuss your QC requirements.

The single-stranded DNA is supplied frozen in endotoxin-free water, TE buffer, or freeze-dried at room temperature. Other delivery options are possible upon request.

1. Spin your tube upon arrival for 5 seconds in a microcentrifuge, to guarantee that your single stranded DNA is at the bottom of the tube.
2. Add water according to the CoA to achieve a concentration of 100 µM.
3. Mix briefly by flicking or vortexing the tube.
4. Incubate overnight at 4 °C or at room temperature for 1 h.
5. Vortex and centrifuge briefly.
6. Verify the final concentration with a nano volume spectrophotometer.

Long-term storage at -20 °C and short-term storage at 4 °C is recommended. Resuspend in nuclease-free water or TE-buffer for best results. The stability of single-stranded DNA is dependent on the buffer as well as the storage temperature.

The gxstrands process, from initial inquiry to final delivery, spans approximately 6 weeks. Turnaround time varies depending on sequence complexity and scale.

For precise information regarding your specific delivery schedule, please contact info@gxstrands.com