Oligonucleotide _ Oligonucleotide Synthesis _ Oligonucleotide Solutions

Oligonucleotides are small molecule biopolymers formed by the connection of multiple nucleotides, usually consisting of less than 20 nucleotides. They are widely used in biological, medical, forensic, and clinical applications, such as as primers and probes for PCR and Sanger sequencing, and as therapeutic agents.

Oligonucleotide synthesis method

The main synthetic methods of oligonucleotides include solid-phase synthesis. The method includes the following steps:

Deprotection: Removal of protective groups on nucleotides using organic solvents and acid solutions.

Coupling: Activated monomers react with deprotected hydroxyl groups to form new phosphoroxy bonds and extend nucleotide chains.

Oxidation: formation of phosphodiester bonds.

Capping: Prevent hydroxyl groups that are not involved in the reaction from continuing to react.

Ammoniolysis: cleavage of oligonucleotides from the solid support.

Purification and ultrafiltration: removal of impurities by chromatography and ultrafiltration.

Oligonucleotide application field

Oligonucleotides are widely used in research and diagnostics, including:

Primers are used for enzymatic reactions such as PCR to initiate the amplification of DNA or RNA.

Probes are used to detect specific DNA or RNA sequences.

Therapeutic drugs, such as antisense oligonucleotides (ASO) and small interfering RNA (siRNA), are used for gene silencing and disease treatment.

Challenges and Solutions in Oligonucleotide Production

In the oligonucleotide production process, the main challenges include explosion protection and water removal requirements. Due to the use of large amounts of organic solvents, production facilities need to be specially designed to prevent explosion risks. In addition, strict control of the moisture content of solvents and monomers is required during the synthesis process. Class 3A molecular sieves are usually used for water removal treatment.