What is Oligonucleotide Synthesis?
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- Опубликовано: 24 ноя 2020
- Oligonucleotides can be formed either enzymatically through cleavage of larger biomolecules or through targeted chemical synthesis. In the latter case, organonucleotide synthesis is the chemical process by which nucleoside phosphoramidites - a key monomeric element of oligonucleotides - are synthesized. Nucleoside phosphoramidites (or nucleosides in shorthand) themselves are derivatives of natural or synthetic nucleosides.
Nucleotides, therefore, are compound molecules consisting of a covalently linked nucleoside and a phosphate group. Nucleotides are linked in a specific sequence to form the desired product. Thus, oligonucleotides consist of short segments of nucleic acids that are linked together forming single-chain biological polymers (“-mers”). Since most oligonucleotides (or oligos in shorthand) typically consist of up to 20 linked nucleotides (though more are possible), they can be thought of as small biopharmaceutical molecules.
Much like other pharmaceutically-relevant small molecules, oligonucleotide synthesis requires careful control with respect to temperature, pH, quality of substrates, etc. Because of the typical methods used in oligonucleotide synthesis, the longer and more complicated the nucleotide sequence, the more difficult it is to meet yield, purity, and cost objectives. Process Analytical Technologies (PATs) are useful in achieving these goals.
Learn more about oligonucleotide synthesis: www.mt.com/global/en/home/app...
#oligonucleotides #oligonucleotidesynthesis #oligo 
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I've been working for an oligonucleotide synthesis company for seven years and I've never heard the chemistry of what we do explained so clearly and concisely before. Well done!
can you tell me what you actually do without the chemistry analysis? I mean what liquids do you mix together and what temperatures and mixing you use to get the results? where can I find out about this? also how did you get a job for such a company?
It's easy to understand, thank you!
Learn more about Oligonucleotide Synthesis: bit.ly/3WyuJVX
Todo un trabajo espectacular, sintetice por primera vez en Colombia y de forma manual cadenas de ADN por el método del fosforamidito . Mi tesis de grado fue exaltada y me gradué con honores como Químico Farmacéutico. Monte un microlaboratorio
Vengo a Colombia en 3 semanas y me encantaría reunirme con usted y hablar con usted sobre esto, estoy muy interesado en la ingeniería genética. Y la síntesis de oligonucleótidos es uno de los pasos más importantes.
Como cuanto sale en dinero una cadena de 10 oligonucleotidos?
Thank you..the was really helpful ✨
Where can I see the lab demonstration of this process?
Unfortunately, we do not have a lab demonstration video of this process just yet. However, you can go to this page and "Schedule a Free eDemo" to request what you would like to see! bit.ly/3WyuJVX
i am so sorry for this dumb question but im late at understanding clearly. so what happens is that the dna unwinds and the template starts all the transfer reactions and nucleotides are added until the length is reached and then that long template cleaves into two which gives us our new dna... am i right? so the dna template doesnt start off the scratch ut continues to grow on the template and then cleaves into two. did i get that right????
In the method shown here there isn't a template. The synthesis starts with a single base and then attaches them in a chain one at a time. The sequence is determined before hand and controlled by only allowing one monomer at a time to couple onto the sequence.
@@jordanvozenilek1092 your explaination made it more clear. Thanks
Thank you ......How can I bind Zinc Oxide nanoparticles to an amin linked oligotargeter?
Thank you for your question. While we do not have the specific experience for design of zinc oxide synthetic conjugation chemistry, we would suggest that you reference articles such as the example cited here. While this addresses the chemistry and method for conjugation of antibacterial species, the conjugation process may share similarities for your chemistry. We recommend that you still confirm the reaction scheme with an experienced synthetic chemists/biochemist. To follow reaction kinetics in real-time, the FTIR method described in this paper could be performed by a probe or flow-cell based in situ FTIR with Attenuated Total Reflectance (ATR).
Akbar, N., Aslam, Z., Siddiqui, R., Shah, M. R., & Khan, N. A. (2021). Zinc oxide nanoparticles conjugated with clinically-approved medicines as potential antibacterial molecules. AMB Express, 11(1). doi.org/10.1186/s13568-021-01261-1
I don't get how we synthesize exact oligonucleotide which i want .... it's random
The chemical steps in each cycle called "activation", "detritylation (a form of de-blocking)", "coupling", "capping" and "solvent washes" control the product oligonucleotide order and prevent/minimize bad sequences (impurities called sequence failures). In more detail, a target sequence for a product oligonucleotide should be known before synthesis (i.e. the medical disease or biotech purpose being targeted). Then a reverse-complementary (anti-sense) sequence is digitally designed. Next, an oligonucleotide synthesizer or other production rig will attach multiple stock solutions called phosphoramidites (amidites for short) connected in known positions on the synthesizer. Each amidite has a different nucleoside base from the others (i.e. A, C, G, T or synthetic versions). As the synthesizer reads the designed digital sequence, the rig will added only one of the amidites at a time into the reactor or column. Let us assume our sequence is "Linker-ACCTTAGG"... and we just added another volume of 'A' into the reactor. A specific part of the A-amidite will react with the 'tail' of the last 'G' on the sequence at the 5-prime carbon. After the amidite reacts to (is coupled to) the previous amidite in the sequence, 'G', there may be residual or unreacted A-amidite left over. So we now have "Linker-ACCTTAGGA" plus some extra 'free-A'. We need to remove this excess by washing out the reactor or column with an appropriate solvent. Because chemistry is not 100% efficient, we might have 99 sequences that reacted correctly to become "Linker-ACCTTAGGA", but we had 1 sequence fail to react and is stuck as "Linker-ACCTTAGG". In order to prevent that 1 sequence from becoming an impurity known as a sequence failure, we will chemically "cap" the end to prevent it from becoming "Linker-ACCTTAGGC" in the next reaction cycle when we have added C-amidite and the other 99 sequences have reacted to become "Linker-ACCTTAGGAC".
@@MettlerToledoAC Thank you so much for the video and detailed comments! It really helped me to do last minute study for my midsems