Solid Phase Oligonucleotide Synthesis

With a state-of-the-art molecular biology platform, BOC Sciences' experienced oligonucleotide R&D and production team provides you with high-quality, customized, large-scale oligonucleotide synthesis services. We are equipped with state-of-the-art equipment and technology to synthesize DNA and RNA oligonucleotides in the mg to kg range.

Fig 1. Schematic of the solid-phase synthesis of PS oligonucleotides. (Yang et al., 2018)

What is Solid Phase Oligonucleotide Synthesis?

Oligonucleotides can be synthesized by solid-phase phosphoramidite chemistry, and the synthesis method is the solid-phase phosphoramidite method (referred to as solid-phase synthesis). The synthesis vehicle plays an important role in the solid phase synthesis reaction. Commonly used carriers include controllable microporous glass (CPG) and polystyrene (PS). CPG carriers have the advantages of good mechanical properties, insolubility, and controllable pore size. PS carriers have the advantage of high loading (350 μmol/g) and can be applied as a large-scale synthesis of oligonucleotide carriers. Solid-phase synthesis of oligonucleotides is an important technique to synthesize short nucleic acid sequences by adding nucleotide units one by one on a solid-phase support. With high purity, high yields and a wide range of applications, this synthesis method is important for nucleic acid-related work in research and applications.

Advantages of Solid Phase Synthesis of Oligonucleotide

Solid phase synthesis is carried out on a solid support between filters in a column that allows all reagents and solvents to pass freely. Solid phase synthesis offers many advantages over solution synthesi.

• Large excesses of solution phase reagents are available to drive the reaction to rapid completion.
• Impurities and excess reagents are washed away and purification is not required after each step.
• The process lends itself to automation on a computer-controlled solid phase synthesizer.

Steps in Solid Phase Synthesis

Each cycle of solid phase synthesis consists of four main steps: deprotection, coupling, oxidation, and capping. Each cycle attaches a new nucleotide until the oligonucleotide chain is extended to the desired length. The oligonucleotide is then treated with ammonia to cut the oligonucleotide from the solid phase carrier and remove the protective groups, followed by a series of purification, desalting and lyophilization steps to obtain the final purified oligonucleotide.

Solid Phase Oligonucleotide Synthesis Service

• Customized Oligonucleotide Production

  
  Classical solid-phase chemical synthesis of oligonucleotides usually involves the use of polymers or specialized glass beads that are not affected by the reaction conditions. The purpose of the beads is to act as a platform for covalent binding of substrate molecules, which then act through the chemical reaction. Solid-phase synthesis is effective in producing purified products quickly because impurities and unreacted materials are washed away during the various steps of the synthesis.

• Automated Synthesis of Oligonucleotides

  
  Solid-phase oligonucleotide synthesis can now be automated using computer-controlled instrumentation and is technically implemented in column, multiwell plate and array formats. This column format is well suited for research and large-scale applications that do not require high throughput. The multiwell plate format is designed for small-scale, high-throughput synthesis to meet the growing demand for synthetic oligonucleotides in industry and academia.

• Modification of Oligonucleotides

  
  Unmodified oligonucleotides are susceptible to nuclease degradation upon entry into the body and show unfavorable cellular uptake and biodistribution. Positions on oligonucleotides that are commonly modified include the addition of phosphate backbones (e.g., phosphorothioate bond PS, etc.), ribose (e.g., 2'-OMe, 2'-F, and nucleic acid sugar LNA, etc.), pyrimidine bases (e.g., 5'-methylcytosine, etc.), and targeting ligands (e.g., GalNAc, etc.).

• Quality Analysis of Oligonucleotides

  
  Currently, BOC Sciences provides oligonucleotide quality analysis methods such as HPLC, LC-MS, ELISA, RT-qPCR, and CGE.

Advantages of BOC Sciences' Oligonucleotide Synthesis Service

• More synthesis scale - Synthesis volume can meet customers' needs at different stages
• Higher purity and lower toxicity - Purity can reach over 90%, with better purification process and lower cytotoxicity.
• More modification types - A variety of chemical modifications and fluorescent labeling
• Customized service - Senior R&D technical support team provides customized service.

BOC Sciences has the technology and capability in solid-phase synthesis of oligonucleotides to efficiently meet customers' requirements within the delivery time. If you are interested in our services, please feel free to contact us.

Reference

1. Yang J, et al. Solid-Phase Synthesis of Phosphorothioate Oligonucleotides Using Sulfurization Byproducts for in Situ Capping[J]. The Journal of Organic Chemistry, 2018, 83(19): 11577-11585.

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Solid Phase Oligonucleotide Synthesis

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