Click chemistry allows for multiple labeling of polymerized DNA strands via incorporation of Alkyne-dNTPs (triphosphates) followed by post-synthetic click reactions at a so far unmatched efficiency level. Want to learn more about it?
Since 2010, Click Chemistry has become a hot topic in oligonucleotide labeling, now reaching a peak of interest with the recent Nobel Prize in Chemistry 2022 awarded to scientists Carolyn R. Bertozzi, Morten Meldal and K. Barry Sharpless for their development of click chemistry and bio-orthogonal chemistry.
Always striving to be at the forefront of technical improvements to better serve our customers, metabion has secured a worldwide license for employing this interesting chemistry in the production of modified oligonucleotides.
Initially, the term "click chemistry" was coined by K. Barry Sharpless in 1998, and was fully described by Sharpless, Hartmuth C. Kolb and M.G. Finn of the Scripps Research Institute in 2001. It is a versatile technique that can be used for the synthesis of a variety of conjugates. Virtually it can be applied to any biomolecules. With respect to oligonucleotide labeling, click chemistry allows multiple labeling of polymerized DNA strands via incorporation of Alkyne-dNTPs (triphosphates) followed by post-synthetic click reactions at a level of efficiency unmatched to date.
Want to learn more about the concept of click chemistry? Visit our Knowledge Hub!
Which applications are possible with click chemistry?
Extended means of (multiple) labeling of biomolecules such as Nucleic Acids (DNA/RNA), peptides, proteins, and antibodies will have a vigorous effect on the following areas (making no claim to be exhaustive):
- Fluorescence In Situ Hybridization (FISH)
- qPCR-Assays
- DNA/RNA Microarrays;
- Bioconjugation to Nanoparticles
- Fluorescence Microscopy
- Electrophoretic Mobility Shift Assay (EMSA)
- Fluorescence Activated Cell Sorting (FACS)
- Flow Cytometry (FCM), Cell Tracking and Cell-based Assays
- Fluorescence Correlation Spectroscopy (FCS)
- …
What we offer
1. “ready-to-use/fully labelled” custom oligonucleotides
Having successfully introduced this additional labelling procedure for synthetic Nucleic Acids and other biomolecules into our custom synthesis operations, we are now able to offer significantly increased flexibility in terms of:
- Multiple labeling of oligonucleotides (2 or more modifications);
- Producing modified oligonucleotides exceeding a critical sequence length.
Send your requests to info@metabion.com. We will come back to you with solutions and/or proposals that will surprise you.
2. “ready-to-click-it-yourself” custom oligonucleotides
Moreover, we can provide you with alkyne-modified
oligonucleotides to be labelled at the oligo end by “clicking” the desired
“azide-modification”. For more information, see our full list of
Click-modifications below.
There are 2 ways to introduce Alkyne groups to the 5´end of the oligo:
- You can introduce an Alxyl moiety, which is sequence independent and allows the addition of an Alkyne group via a C6 (hexynyl)- linker to the phosphate of the 5´ nucleoside.
- If your oligo sequence carries a C or T at the 5´ end, you can introduce an Alkyne-activation site via replacing the C/T with AldC/ AldU, respectively (sequence dependent). AldU and AldC carry a C8-Alkyne motif at the C5 position of the respective Pyrimidine structure of either dU (replacing dT) or dC.
For internal Alkynylation, you can either choose AldU or AldC to replace dT and/or dC within the oligo sequence.
For Alkyne-activation of the 3´ end, again, there are two options:
- You can introduce an Alrol moiety, which is sequence independent and connects your oligo with an Alkyne group via a serinol-linker to at the 3´ OH of the 3´ nucleoside (thus producing a 3´OH-blocking);
- If your oligo sequence carries a C or T at the 3´ end, you can introduce an Alkyne-activation site via replacing the dC/ dT with AldC/ AldU, respectively (sequence dependent). AldU and AldC carry a C8-Alkyne motif at the C5 position of the respective Pyrimidine structure of either dU (replacing dT) or dC. This way, you can introduce a 3’ Alkyne-activation site without blocking the 3´OH end.
For detailed information about how to order, visit our FAQs page.
3. “modimerization” solutions
In line with above´s “click it yourself” approach, we can also supply you with:
- Alkyne Nucleotides (triphosphates) to be incorporated into DNA during polymerization reactions (PCR, reverse transcription, nick-translation, etc.). This prepares the produced DNA strands for post-synthetic “click” conjugations to azide-activated labels – a very elegant and efficient way of “modimerizing” DNA strands;
- Azide Modifiers and additional “click reagents”. Azide-activated molecules can be used to modify alkynylated DNA/RNA sequences and other alkynylated biomolecules.
For detailed information about how to order, visit our FAQs page.
How to order
Use our Web Order Portal: Order now
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E-mail
Please send an e-mail to mi@mymetabion.com specifying your order as well as your shipping and billing address, and any other information which might be required like PO number, VAT number (foreign country EU countries, beside Germany), etc.