Production

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quality – flexibility – traceability – reliability

Diagnostic

Our diagnostic oligonucleotide synthesis service has been certified according to ISO 13485:2003 since 2011. Production, purification, final-fill and shipping is undertaken in dedicated labs under controlled conditions and meet the elevated quality standards required for products used in clinical or molecular diagnostic applications.

Specifications of purification and QC methods, dedicated columns, Certificate of Analysis and quality standards can all be defined by the customer themself.

Special synthesis
Internal modifications, 5´Quencher – 3´Reporter, Inosin/Wobbles and many more synthesis options available

Purification options
Purification of primers and probes differs according to sequence and dyes. Typically we use RP/IX-HPLC.
Please enquire for more purification options.
Dedicated columns for HPLC-purification are available on request.

Turnaround time
Depending on the size of the order we need at least 5 working days.

QC options
Maldi-MS, ESI-MS, HPLC and PAGE
customized „Certificate of Analysis“ on request

Overview Reporter-Quencher

A unique service for randomized oligos

Trimer – Technology

The use of 2′-deoxynucleoside trimer phosphoramidites is a particularly efficient way to approach oligonucleotide-directed mutagenesis.
Of the 64 possible combinations of codons, only 20 codons are required to cover the 20 amino acids.
Unlike other methods of mutagenesis, libraries built of Trimer-oligonucleotides lead to no codon bias, no production of stop codons and custom mixes allow for control of codon abundance making them one of the most efficient tools to explore sequence space in protein regions that are important for function – even in nonsaturating conditions.
All available trimers which are listed in the link below have the protection scheme described by Kayushin et al (2-4).
The reaction factor of each new trimer-phosphoramidite lot will be determined by Ella Biotech before used in synthesis. The reaction factor is critical since the trimers will likely be mixed and they have differing reactivity in the coupling reaction.
With this method we can guarantee a correct distribution of the trimer-codons in the mix.

Application

  • library construction for protein mutagenesis using 2′-deoxynucleoside trimer phosphoramidites

Advantages

  • no codon bias
  • no frame-shifts
  • high diversity of sequences against Standard NNK or NNS libraries
  • no stop codons

Quality and Service

  • We guarantee a variance in the allocation of each trimer in a 19-trimer mix of 2-9%. This is based on statistics of sequencing 300 clones!
  • We produce our degenerated oligos to 90-95% purity, based on 70mers with 4 to 8 couplings of trimer mixes. For purification we use HPLC and PAGE. The HPLC-method we use has been developed specifically for the purification of trimer-oligonucleotides which have given excellent results in successful trimer-projects.
  • All Trimer-codons will be quality checked by NGS before used in synthesis
  • Several modifications available for Trimer-Oligonucleotides (e.g. Biotin, Phos, Spacer etc.)
  • Special method used for library synthesis with length polymorphism
  • synthesis follows our quality standards according to ISO9001
  • further trimer-codons (custom codons) are available which are not listed in the overview – please enquire
  • high quality amplification primers will be provided free of charge
  • guaranteed yield: 3-5 nmol

Turnaround time
Depending on the size of the order we need at least 10 working days – please enquire

Overview Reporter-Quencher

fast – reliable – customized

Standard Synthesis

We can offer you the full range of DNA Oligonucleotides. In tubes or plate-size, customized tubes, all common modifications, Duplex-DNA, normalization as well as aliquoting service.

Our DNA-Oligonucleotides are available deprotected and desalted or purified by HPLC or PAGE. Standard scales are available in the list. Please contact us if you need higher scales or customized yields.

Custom DNA Oligos in tubes

  • length up to 200 bases
  • standard scale and customized final yields
  • wide range of modifications available
  • final QC by e.g. MALDI-TOF, ESI-MS, Page – on request

Custom DNA Oligos in 96-Well Plates

  • length up to 60 bases
  • desalted quality
  • normalizing service included

Duplex Oligonucleotides

  • set-up costs: EUR 50,00/strand (please enquire for more information)
  • additional HPLC after annealing: EUR 50,00/duplex

Additional service

  • normalizing, aliquoting, customized wobble-mixes, customized QC

Turnaround Time

  • non-modified oligos: 1 day
  • non-modified HPLC oligos: 2 days
  • modified Oligos: 2-3 days

Scales and Yields

Modifications

OD-nmol Conversion

specific – robust – stable – sensitive

LNA-OLIGONUCLEOTIDE Production

Locked Nucleic Acids (LNA) are a class of synthetic nucleic acid analogues that contain a “locked” bicyclic sugar moiety. This linkage between the 2’-O- and the 4’-C-position locks the ribose in the 3´-endo conformation (see Figure 1) which leads to the characteristic structure of A-form RNA and forms exclusively A-type duplexes.

Compared to traditional DNA or RNA oligonucleotides, the use of LNA modified oligos exhibit enhanced hybridization affinity towards complementary DNA or RNA.1) LNA oligos can be synthesized using conventional phosphoramidite-chemistry to increase the Tm by 3-8°C for each substituted nucleotide at simultaneously shorter length compared to traditional DNA or RNA oligonucleotides.2) This helps e.g. to detect shorter or very similar target sequences.

Structures of DNA, RNA and LNA with its C3′-endo, Northern, A-form conformer

  • Finetuning of melting temperature and Tm normalization
  • Enhanced binding affinity at simultaneously shorter length
  • Improved signal to noise ratio in qPCR Assays
  • Increased target specificity and high sensitivity compared to traditional DNA or RNA probes
  • Enhanced single nucleotide mismatch detection
  • High in-vitro and in-vivo stability due to increased endo- and exonuclease resistance
  • Easy cell entering of antisense LNA oligonucleotides due to negatively charged backbone (combined with cationic transfection agents)
  • Potent & nontoxic building blocks in antisense drug development3)
  • Gene silencing with antisense LNA oligonucleotides
  • qPCR/PCR applications and Multiplexing
  • Methylation analysis
  • Aptamers
  • Allele discrimination
  • Gene expression analysis
  • Microarray analysis
  • Avoid stretches of more than 4 LNA bases, except when very short (9-10 nt) oligonucleotides are designed
  • 3 +G and 3 +C stretches should be also avoided
  • If just one LNA base is positioned at the 5´end, there won´t be any influence on Tm
  • Keep the GC-content between 30-60 %
  • LNA bases should not be positioned at the 3´end or near the 3´terminus
  • Not more than 15 LNA bases should be incorporated because it increases the risk of self-hybridization
  • Single LNA base increases Tm by 2-6°C (DNA) and 3-9°C (RNA) upon binding
  • All standard scales and customized final yields are available
  • LNA should be marked in the sequence with + (e.g. +A, +C, +G, +T)
  • Full compatibility for all additional modifications in our DNA/RNA Portfolio
  • For any further information please enquire

fast – reliable – customized – high quality standards

RNA

We can offer you the full range of RNA Oligonucleotides. Standard-RNA, modified, O-Methyl, DNA/RNA Chimera as well as Duplex-RNA.
Our RNA-Production follows our high quality standards and all produced RNA Oligos will be QC´d by Maldi or according to the customer’s request.

  • Custom RNA Oligos
    • length up to 60 bases
    • standard scale and customized final yields
    • wide range of 5´and 3´modification available – please enquire

    Custom 2´-O-Methyl RNA Oligos

    • length up to 60 bases
    • standard scale and customized final yields
    • wide range of 5´and 3´modification available – please enquire

    Custom DNA/RNA Chimera

    • length up to 60 bases
    • standard scale and customized final yields
    • wide range of 5´and 3´modification available – please enquire

    Duplex RNA

    • setup-costs for annealing: EUR 50,00/single-strand
    • additional HPLC after annealing: EUR 50,00/duplex

    Long-RNA

    • length up to 150 bases
    • wide range of 5´and 3´modification available
    • please enquire

    Additional service

    • normalizing, aliquoting, customized wobble-mixes, customized QC

    Turnaround time

    • unmodified RNA: ~ 5 working days
    • modified and Duplex RNA: ~5-7 working days
Scales and Yields

Large Scale – Oligonucleotide Production
high quality – competitive service

Large scale

Let us help you determine the best large-scale solutions for your projects, so you can focus on generating informative results instead of spending time and money on troubleshooting.
Our production capacity and synthesis expertise allow us to manufacture highly complex oligos for in vivo, high-throughput, or commercial applications. If needed, we can supply you with grams of highly pure siRNAs/DsiRNAs, aptamers, antisense oligos, ribozymes, miRNAs, decoy oligos, next generation sequencing products, and qPCR primers and probe sets.

  • final yields up to several 100µmol final yield (purity guarantees and synthesis options may vary depending on scale)
  • standard and modified bases
  • almost any chemical modification, fluorophore, or dark quencher
  • custom oligo mixtures available
  • flexible ordering methods (e.g., use the units that work best for your projects)
  • customer-dedicated purification columns available
  • choice of aliquoting and vialing formats
  • independently verified product compliance and release prior to shipment (Certificates of Analysis)
  • hardcopy of analytical QC documentation provided

We need at least 10 working days for standard oligos depending on project size

Ella Biotech GmbH | Am Kugelfang 39, D-82256 Fürstenfeldbruck | info@ellabiotech.com | +49 8141 36636-0

ISO 13485

The International Organization for Standardization (ISO) is a voluntary, non-governmental organization that provides unified standards for many different areas of industrial activity. The ISO 13485 designation is the standard for manufacturers of medical diagnostics. While similar to GMP in many respects, holding an ISO 13485 certification is not a legal requirement, but is accepted as an international standard. Manufacturers are continually audited and must be recertified on a regular basis. The regular audits ensure that the reported standards of manufacturing are maintained and encourage continuous improvements.
As of 2010 we have observed an increasing demand for high quality oligos for use in In Vitro Diagnostics. Therefore, Ella Biotech determined to obtain the ISO 13485 certification to provide proof that we have a system in place to validate and provide traceability for oligonucleotide manufacturing history during the production process. In May 2011 we were successfully certified by DQS under certificate registration no. “345862 MP23SCC” for “Development, Production and Distribution of Biopolymers for the use in In-Vitro Diagnostics”.
Together with our customers in human diagnostics, we define the oligo characteristics that will work best for their diagnostic applications. We provide guidance on process scale-up for deliverable yields of a few nmoles up to multiple grams. We also boast supplementary services such as dedicated purification columns, aliquoting and other custom packaging, and oligo mixture formulation.

ISO 9001

ISO 9001 is one of the most widely used management tools in the world today. It ensures that the company meet the needs of customers and other stakeholders while meeting statutory and regulatory requirements related to a product or service. Our Quality Management System is certified by DQS under certificate registration no. “345862 QM08”. The certification applies to following activities: “Development, Production and Distribution of Biopolymers”.