We provide custom primers designed for precision in research and diagnostics. Offering a wide array of modifications like duplex stabilizers and low bias degenerations in conjunction with highly optimized synthesis and purification methods, our primers ensure high performance and reliability for applications like qPCR and sequencing.

In addition, to minimize the risk of contamination, our diagnostic oligos are manufactured in a separate, dedicated laboratory. This ensures that they meet the highest standards of purity and reliability, giving you confidence in their accuracy for diagnostic applications.

Length

We offer custom-made primers from 8 to 40 nucleotides in length, manufactured with a high degree of fidelity for reliable performance.
While we generally don’t produce primers over 40 nt for diagnostics to minimise the risk of contamination, we are happy to discuss special cases – for other lengths or special requirements, please contact us for a tailored solution.

Scales and Yields

We begin primer synthesis at a scale of 40 or 200 nmol. The final yield is influenced by the desired level of purity, as more stringent purification procedures will reduce the yield.

Modifications

Optimizing primers with specialized modifications has been shown to enhance PCR performance by improving detection sensitivity, specificity, and stability. We offer a variety of modification options that provide greater flexibility in oligonucleotide design. This ensures that primers perform optimally in PCR-based diagnostic and research applications.

There is a comprehensive list of all the available modification options. : List of available modifications

Duplex Stabilization

Duplex stabilizing modifications have been shown to enhance primer stability and performance by increasing thermal stability, structural integrity, hybridization efficiency, and binding affinity. These improvements are particularly valuable in PCR, qPCR, and other amplification techniques that require precise and stable target binding. By incorporating these modifications, assays achieve greater accuracy, sensitivity, and specificity, ultimately improving overall performance and reliability in applications.

* Values may vary depending on your oligo design.

RNA and DNA/RNA Chimeras

Single or multiple RNA bases and/or RNA analogue bases such as 2’O-Methyl, 2’Fluoro, and 2’O-Methoxyethyl (MOE) have been shown to improve the stability and binding affinity of DNA and RNA targets. These modifications can reduce mismatch tolerance, making them ideal for many high-specificity applications.

We offer the synthesis of chimeric DNA/RNA primers for specific qPCR applications to take advantage of the inherent positive effects of chimeric qPCR primers.

DNA/RNA Chimeras: 2’O-Methyl, 2′ Fluoro, 2’O-Methoxyethyl(MOE), RNA and LNA.

Degenerate Bases

In oligonucleotide synthesis, the deliberate incorporation of degenerate bases (or „wobbles“) at specific positions within an oligonucleotide sequence is referred to as „degeneration“. The use of such degenerate bases allows the synthesis of oligonucleotides with mixed or variable sequence at specific positions. This technique is essential for applications such as primer libraries for unknown templates, cloning diversity, site-directed mutagenesis, and DNA amplification based on protein sequences or homologous regions across species. 

At Ella Biotech, we offer a range of standard wobbles for different combinations: for example, „K“ represents a mixture of Guanine (G) and Thymine (T), „M“ combines Adenine (A) and Cytosine (C), and „R“ includes Adenine (A) and Guanine (G). For more complex applications, wider degeneracy options such as „N“ (A/G/C/T) can be used.

The universal codes for wobbles:
K (G/T), M (A/C), R (A/G), S (G/C), W (A/T), Y (C/T), B (G/C/T), D (A/G/T), H (A/C/T), V (A/G/C), N (A/G/C/T)

In addition, custom wobble compositions, such as 80% Adenine (A) and 20% Guanine (G), allow for customized nucleotide ratios to meet specific experimental needs.

These wobbles are also compatible with alternative backbones such as Locked Nucleic Acids (LNA) and 2′-O-Methyl RNA, providing researchers with the flexibility to incorporate degeneration modifications into a variety of oligonucleotide chemistries.

In addition to standard wobbles, we also offer degenerate bases designed to increase oligonucleotide flexibility by pairing with multiple bases. For example, the degenerate base dK can pair with Cytosine (C) and Thymine (T), while dP complements Adenine (A) and Guanine (G). Another versatile option is Inosine (dI), which pairs with all four bases (A, G, C, and T), making it ideal for applications requiring base pairing versatility.

We also provide abasic positions such as dSpacer, which leaves a gap in the sequence and can be used to mimic natural DNA damage or to study structural changes in oligos. These specialized options, along with our selection of wobbles, provide scientists with greater control over sequence flexibility and target binding in complex experimental designs.

Purification

Depending on the application, you can request the purity level of your primers. Higher quality often requires more stringent purification steps. However, this can result in lower yields for a given synthesis scale and sequence length due to the removal of impurities and shorter by-products.

For IVD oligos longer than 25 bases, we recommend RP-HPLC purification to ensure maximum reproducibility and reliability of results.

You can find more information about Purification Options in our Resource section.

Delivery Form and Packaging

The oligo products are available in multiple packaging formats, including single tubes, multi-well plates, and bulk options, with concentrations, solutions, and aliquot options to suit your requirements. Custom vial labeling is also available upon request.

* Possible concentration may depend on scale/yield since low yields at high concentrations will lead to extremely low volumes
   which are very hard to handle e.g., 5 nmol at 5 mM would lead to a volume of 1 µL.

Quality and Quality Control (QC)

We ensure the highest quality of our synthesized oligonucleotides by conducting rigorous quality control tests on each oligonucleotide. All oligos are manufactured under ISO 13485 standards to ensure exceptional purity and reliability.

For detailed information please refer to Quality Control page in our Resource section.

Additional Services

Prepaid Oligo Service: You can set up a credit account to make your purchases easier by paying by direct debit from your prepaid credit account.

For quality assurance purposes, we can provide comprehensive Certificate of Analysis (CoA) documentation upon request.

Production Time and Shipment

The time required for production of custom oligos varies based on the complexity of the modifications, the yield, and the order volume. Probes with complex modifications typically require more time than standard primers, and larger orders or higher yields can extend the timeline. We strive to meet estimated delivery times and provide updates along the way.

The production time for primer oligos averages 3 working days, typically ranging from 3 to 5 days depending on the yield/order volume and additional services such as CoAs or custom purification.

We use FedEx for shipment, offering both national and international delivery options. For temperature-sensitive shipments, we provide the option of dry ice or blue ice (gel packs).