Hybrid Oligonucleotides Synthesis

Hybrid oligonucleotide synthesis is a method for synthesizing short DNA / RNA molecules containing natural and modified nucleotides. The process involves incorporating modified nucleotides into the oligonucleotide sequence to enhance its stability, binding affinity, or resistance to nuclease degradation. Hybrid oligonucleotide synthesis has revolutionized the field of nucleic acid chemistry by providing a powerful tool for the design and synthesis of nucleic acid probes and therapeutic agents with enhanced properties and better performance.

How to Synthesize Hybrid Oligonucleotides?

Hybrid oligonucleotides are typically synthesized using solid phase synthesis, where oligonucleotides are constructed stepwise on a solid support. Modified nucleotides are added to the oligonucleotide sequence along with natural nucleotides using standard phosphoramidite chemistry.

The synthesis of hybrid oligonucleotides also involves the incorporation of modified nucleotides, such as phosphorothioate nucleotides or 2'-O-methyl nucleotides, into the oligonucleotide sequence along with the natural nucleotides.

BOC Sciences' Hybrid Oligonucleotides Preparation Services

BOC Sciences can provide customers with hybrid oligonucleotide preparation services, which typically include the synthesis of custom oligonucleotides containing modified nucleotides, as well as purification and characterization of the final product. Our specific oligonucleotide hybridization services include the following:

• Hybrid Oligonucleotides Design

When performing oligonucleotide sequence design, the purpose of the oligonucleotide needs to be determined, such as PCR amplification, gene expression analysis, or target binding. And then make the selection of the target region, if the oligonucleotide is designed for PCR amplification or sequencing, select a specific target region in the DNA or RNA sequence. Determine the optimal length of the oligonucleotide based on the desired application and the characteristics of the target sequence.

Design with care: Tm should also be considered when designing an oligonucleotide to ensure efficient hybridization or amplification and to avoid regions of the target sequence that can form secondary structures, such as hairpins or loops, which can interfere with hybridization or amplification.

• Hybrid Oligonucleotides Synthesis

We can synthesize oligonucleotides using standard solid phase synthesis methods such as phosphoramidite chemistry. In addition, many modifications can be added to the oligonucleotide sequence to alter its properties and function.

Some of the most common modifications include:

(1) Phosphorothioate linkage

In a phosphorothioate linkage, an unbridged oxygen atom in the phosphate group is replaced by a sulfur atom. This modification increases the stability of the oligonucleotide and improves its resistance to nuclease degradation.

(2) 2'-O-methyl nucleotides

2'-O-methyl nucleotides attach a methyl group to the 2'-position of the ribose. This modification increases the stability of the oligonucleotide and improves its binding affinity to complementary RNA targets.

(3) Phosphate-modified nucleotides

Phosphate-modified nucleotides contain a phosphate group that is modified by various chemical groups, such as biotin, fluorescein, or amino groups. These modifications can be used to label oligonucleotides for detection or attachment to solid supports.

(4) Fluorescent Dyes

Fluorescent dyes can be attached to oligonucleotides to label oligonucleotides for various applications such as fluorescence in situ hybridization (FISH) or real-time PCR.

• Cleavage and Deprotection

After synthesis of the hybridizing oligonucleotide is complete, the oligonucleotide is cleaved from the solid support and deprotected to remove any residual protecting groups.

• Hybrid Oligonucleotides Purification

Crude oligonucleotides can be purified using a variety of methods to remove any impurities and ensure that the final product is of the desired purity.

• Hybrid Oligonucleotides Characterization

Purified oligonucleotides can be characterized using various techniques such as UV spectrophotometry or mass spectrometry to confirm their identity and purity.

If you are interested in our services, please do not hesitate to contact us at any time.