Custom Circular Oligonucleotide Synthesis Service

Introduction

Creative Biolabs' Custom Circular Oligonucleotide Synthesis delivers precisely engineered nucleic acids for demanding projects, offering enhanced stability, improved binding affinity, precise sequence control, and high yields. Ideal for gene therapy, diagnostics, DNA nanotechnology, and advanced research, it ensures superior quality and performance to support robust scientific endeavors.

[Discover How We Can Help - Request a Consultation]

Custom Circular Oligonucleotide Synthesis

Oligonucleotides are key in modern biology, but linear ones are limited by nuclease susceptibility and stability/specificity issues. Circular oligonucleotides, single-stranded DNA/RNA with a covalently closed loop, solve this: their closed structure, lacking free 5' and 3' ends, grants strong exonuclease resistance and enhanced stability, plus improved binding affinity/specificity. Scalable methods like rolling-circle amplification enable high-yield, cost-effective production, expanding their use in areas like cancer immunotherapy, in vivo therapeutics, and long-term diagnostics.

Fig.1 Selective amplification of oligonucleotides was carried out using cyclic oligonucleotides.¹

Workflow

Required Starting Materials

• Target Sequence: The complete DNA or RNA sequence of your desired oligonucleotide.
• Desired Modifications: Specific details on any modifications (e.g., fluorophores, biotin, phosphorothioate linkages) and their precise locations.
• Quantity Required: Your target yield, ranging from micrograms to milligrams, to tailor the synthesis scale appropriately.

Synthesis

Our custom circular oligonucleotide synthesis begins with high-fidelity linear precursors (incorporating designed sequences and modifications), which are then cyclized via enzymatic ligation to form phosphodiester bonds. Two key enzymatic strategies underpin this process:

• Permuted Intron-Exon (PIE) Method: Utilizing the self-splicing property of type I introns, linear RNA precursors are cyclized via a specific splicing mechanism. It is suitable for larger fragments and relatively simple to operate, but may introduce exogenous sequences that affect function.
• T4 Ligase Method: Using T4 DNA/RNA ligase, the 5' phosphate and 3' hydroxyl groups of linear oligonucleotides are linked with ATP assistance. A splint sequence needs to be designed to facilitate this process. It offers high ligation efficiency and avoids additional chemical groups, but intermolecular side reactions require control.

Purification

Post-amplification, rigorous purification ensures circular oligonucleotide quality. Creative Biolabs uses advanced anion exchange HPLC to separate desired products from unreacted primers, linear fragments, truncations, and residual enzymes. Its high-resolution isolates highly pure full-length circles, vital for sensitive applications where impurities could disrupt results or efficacy.

Quality Control

Every batch of custom circular oligonucleotides undergoes comprehensive validation to ensure unparalleled quality and integrity. Key QC analyses include Mass Spectrometry (MS) and Polyacrylamide Gel Electrophoresis (PAGE).

Estimated Timeframe

The typical timeframe for this service ranges from 4 to 8 weeks, depending on the complexity of the oligonucleotide sequence, the nature and number of required modifications, and the desired synthesis scale. Our team will provide a more precise estimate after reviewing your project specifications.

What we can offer

[Experience the Creative Biolabs Advantage - Get a Quote Today]

Customer Reviews

"Using Creative Biolabs' Custom Circular Oligonucleotide Synthesis in our research has significantly improved the in vivo stability of our therapeutic RNA constructs, leading to more consistent and prolonged gene expression. The nuclease resistance is truly remarkable compared to linear counterparts, addressing a critical bottleneck in our gene therapy development."

[2 months ago], [J. Smi***]

"The high purity and precise sequence control offered by Creative Biolabs' circular oligonucleotides were critical for our DNA origami projects. We achieved unprecedented folding yields and structural integrity, which was challenging with conventionally synthesized oligos. Their comprehensive QC data, including MS and PAGE, was exceptionally thorough and provided great confidence in our experimental setup."

[1 week ago], [M. Cha***]

"Creative Biolabs' scalable amplification method for circular oligonucleotides has drastically reduced our costs for high-throughput screening applications. We can now obtain nanomole quantities of diverse sequences without compromising on quality or specificity, a significant advantage over traditional methods like PCR, which often yielded less pure and less stable products."

[6 months ago], [A. Pat***]

FAQs

Contact Our Team for More Information and to Discuss Your Project

Creative Biolabs is your trusted partner for high-quality Custom Circular Oligonucleotide Synthesis. Our advanced capabilities and commitment to precision ensure superior, tailored products for your R&D needs, accelerating breakthroughs in therapeutics and DNA nanostructures.

Reference

1. Schmidt, Thorsten L., et al. "Scalable amplification of strand subsets from chip-synthesized oligonucleotide libraries." Nature communications 6.1 (2015): 8634. DOI: 10.1038/ncomms9634. Distributed under Open Access license CC BY 4.0, without modification.