The Crucial Role of Oligonucleotide Phosphorylation

Oligonucleotide phosphorylation entails the addition of phosphate groups to nucleotide units in DNA or RNA strands. This modification holds essential roles in biological and synthetic processes, including DNA replication, RNA transcription, and molecular engineering. DNA and RNA polymerases use phosphorylated oligonucleotides as substrates and primers during amplification methods like PCR. Moreover, these modified molecules contribute significantly to cellular signal transduction pathways and regulatory mechanisms. In synthetic biology, phosphorylated oligonucleotides act as foundational elements for crafting altered nucleic acids with enhanced stability or improved binding affinity. These applications span diagnostics, therapeutics, and molecular research, showcasing the intricate impact of oligonucleotide phosphorylation in diverse scientific domains.

Fig.1 Synthesis of 2′-phosphorylated oligonucleotides. (Novopashina, et al., 2007)

Oligonucleotide-based Phosphorylation Modification Service at CD BioGlyco

Oligonucleotide phosphorylation can occur at the 5' or 3' end of the nucleotide unit, typically involving the attachment of a phosphate group to the sugar moiety. These phosphorylation sites are crucial for various biological processes, including enzymatic reactions like DNA replication, transcription, and translation. Based on different sites' phosphorylation modifications by chemical strategies, CD BioGlyco provides a custom oligonucleotide phosphorylation modification service for clients. We will generate suitable and customized proposals for phosphorylation modification according to clients' needs. Besides phosphorylation modification, CD BioGlyco also provides Nucleoside & Nucleotide Synthesis Services, Custom Oligonucleotide Synthesis Services, and other Oligonucleotide Modification Services.

Fig.2 The flowchart of standard oligonucleotide-based phosphorylation modification. (CD BioGlyco)

• Selection of oligonucleotide sequence: Choose the DNA or RNA sequence that requires phosphorylation modification for enhanced stability or functionality.
• Phosphorylation reaction setup: Prepare the reaction mixture containing the oligonucleotide, appropriate buffer, and phosphorylating agents (such as ATP and kinase enzymes).
• Incubation: Allow the reaction mixture to incubate under controlled conditions, often at specific temperatures and durations.
• Quenching reaction: Halt the phosphorylation reaction using quenching agents or by adjusting the reaction conditions.
• Purification: Purify the phosphorylated oligonucleotide to remove unreacted reagents, enzymes, and byproducts. Common purification methods include column chromatography or HPLC.
• Characterization: Confirm the successful phosphorylation modification using techniques like mass spectrometry or gel electrophoresis.
• Scale-Up: If the modified oligonucleotide shows promising results, scale up the synthesis and modification process to meet the required quantities.

Applications of Oligonucleotide-based Phosphorylation Modification Service

• PCR primer design: Phosphorylated primers improve primer-template binding during PCR, enhancing specificity and amplification efficiency.
• DNA/RNA polymerase substrates: Phosphorylated oligonucleotides serve as substrates for DNA and RNA polymerases in enzymatic processes like DNA replication and transcription.
• Aptamer development: Phosphorylated aptamers offer enhanced stability and binding affinity for specific targets, advancing applications in diagnostics and therapeutics.
• Genetic engineering: Phosphorylation facilitates the creation of recombinant DNA constructs, gene editing, and synthetic biology applications.

Advantages of Us

• Our lab team is highly skilled, collaborative, and dedicated, ensuring efficient and accurate execution of experiments and producing high-quality research outcomes.
• Our service team consistently provides exceptional customer support, offering prompt and helpful assistance to address client needs and inquiries.

CD BioGlyco employed highly skilled researchers to process our custom oligonucleotide-based phosphorylation modification experiments. Please feel free to contact us for more information if you are interested in our service.