Oligonucleotide Synthesis

Oligonucleotide synthesis is the chemical synthesis of relatively short fragments of nucleic acids with defined chemical structure (sequence). The technique is extremely useful in current laboratory practice because it provides a rapid and inexpensive access to custom-made oligonucleotides of the desired sequence. Whereas enzymes synthesize DNA and RNA in a 5' to 3' direction, chemical oligonucleotide synthesis is carried out in the opposite, 3' to 5' direction. Currently, the process is implemented as solid-phase synthesis using phosphoramidite method and phosphoramidite building blocks derived from protected 2'-deoxynucleosides (dA, dC, dG, and T), ribonucleosides (A, C, G, and U), or chemically modified nucleosides, e.g. LNA.

To obtain the desired oligonucleotide, the building blocks are sequentially coupled to the growing oligonucleotide chain in the order required by the sequence of the product (see Synthetic cycle below). The process has been fully automated since the late 1970s. Upon the completion of the chain assembly, the product is released from the solid phase to solution, deprotected, and collected. The occurrence of side reactions sets practical limits for the length of synthetic oligonucleotides (up to about 200 nucleotide residues) because the number of errors accumulates with the length of the oligonucleotide being synthesized. Products are often isolated by high-performance liquid chromatography (HPLC) to obtain the desired oligonucleotides in high purity. Typically, synthetic oligonucleotides are single-stranded DNA or RNA molecules around 15–25 bases in length.

Oligonucleotides find a variety of applications in molecular biology and medicine. They are most commonly used as antisense oligonucleotides, small interfering RNA, primers for DNA sequencing and amplification, probes for detecting complementary DNA or RNA via molecular hybridization, tools for the targeted introduction of mutations and restriction sites, and for the synthesis of artificial genes.

Read more about Oligonucleotide SynthesisHistory, Post-synthetic Processing, Characterization, See Also, External Links

Other articles related to "oligonucleotide synthesis, oligonucleotides, oligonucleotide":

Oligonucleotide Synthesis - External Links
... Oligonucleotide calculator – Bio-Synthesis. ...
Oligonucleotide Synthesis - Synthesis By The Phosphoramidite Method - Building Blocks - Nucleoside Phosphoramidites
... analogs are insufficiently reactive to afford an expedite synthetic preparation of oligonucleotides in high yields ... Upon the completion of the oligonucleotide chain assembly, all the protecting groups are removed to yield the desired oligonucleotides ... acetonitrile, the solvent commonly used in oligonucleotide synthesis ...

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