T1 - Enzymatic synthesis of oligonucleotides of defined sequence

There are two ways of ordering oligos with Eton, one is for oligos only and the other is for oligos(primers) that will be used in a DNA sequencing order.

These groups are removed after synthesis of the oligonucleotide is complete, during deprotection.

In general, the longer the oligo, the greater the probability of side reactions during oligo synthesis, along with higher chances to incur into incomplete deprotection. Potential sources of side reactions causing failure products are depurination (which mainly affects the base A) and formation of secondary structures due to the oligos’ sequence. There is no way to completely exclude these effects! However, metabion tries to minimise these failures by continuously optimising synthesis as well as purification protocols!

Our custom DNA portfolio features

These groups are removed after synthesis of the oligomer is complete, during deprotection.

OPC® is an abbreviation for “oligonucleotide purification cartridge”. This purification method consists of a reverse-phase (RP) chromatography, which separates complete, newly synthesized oligonucleotides from failure sequences, by-products, and other impurities. Specifically, the OPC® cartridge binds the dimithyltrityl (DMT) of the trityl-on oligonucleotide (complete, newly synthesized) from non-DMT bearing molecules. This techniques performs best for oligonucleotides up to 40 nucleotides in length.

Custom Oligonucleotide Synthesis

We run two different types of Mass Spectrometry (MS) instruments in order to cope best with quality and quantity/throughput issues determined by the specifications of the respective oligo/analyte. While each instrument type precisely characterizes oligonucleotides in terms of composition through direct molecular weight measurement, their field of application is diligently adjusted to suitability considerations.

PCR, and the synthesis of artificial genes.

ESI-ToF is less efficient in terms of throughput but perfectly compensates for resolution issues with long oligos as well as for a potential detrimental laser impact on labile/photosensitive modifications – thus being a "natural" complement to MALDI-ToF analysis.

A primer is a specific type of oligo

Metabion is dedicated to reliably deliver high quality products. While every production step is performed in light of achieving best quality, the product is released only if it passes our final inspection. Mass Spectrometry has become the state-of-the-art technology for verifying the integrity of oligonucleotides, and metabion has been the first custom oligo house who introduced routine mass checks into its operations. Each and every oligo is characterized by either MALDI- or ESI-ToF and stringent release criteria are applied.

What is an oligo- or oligonucleotide? - Bio-Synthesis Inc

Increase the phosphoramiditeconcentration to enhance the coupling efficiency--e.g., use a concentrationof 50 mg/ml (double the normal concentration; 20-fold molar excess overthe synthetic polynucleotide chain) for longer sequences.

What is an oligo- or oligonucleotide

In addition to what above advised, we recommend that you minimize the exposure of modified oligonucleotides– especially those fluorescently labelled - to light, to avoid any bleaching effect.
Moreover, we recommend storing dye-labelled oligos highly concentrated and not in working dilutions, if you are not planning to use them within 24 hours. The higher the dilution factor, the faster the fluorescent activity fades away. Therefore, try to store highly concentrated aliquots frozen, thaw them only once, dilute them just before use and store the aliquots at 4°C in the dark.