Custom oligonucleotide synthesis has become a cornerstone in molecular biology, enabling researchers to obtain specific DNA or RNA sequences tailored to their experimental needs.
This precision facilitates advancements in various fields, including genetic engineering, diagnostics, and therapeutic development.
Methods of Oligonucleotide Synthesis
The phosphoramidite method stands as the most widely adopted technique for synthesizing DNA oligonucleotides. This approach involves the sequential addition of nucleotides to a growing chain, anchored to a solid-phase support. Each nucleotide addition is carefully controlled, ensuring high fidelity in the resulting sequence. Alternative methods, such as the phosphotriester and H-phosphonate techniques, also exist but are less commonly employed due to their complexity and lower efficiency.
Applications and Modifications
Custom oligonucleotides serve a multitude of applications. In polymerase chain reactions (PCR), they function as primers, initiating DNA amplification. In sequencing technologies, they act as probes, binding to specific sequences to facilitate detection. Moreover, in gene editing,specially designed oligos guide nucleases to precise genomic locations, enabling targeted modifications.
To enhance functionality, oligonucleotides can be modified in various ways. Common modifications include the addition of fluorescent dyes for imaging purposes, incorporation of biotin for affinity purification, and attachment of quenchers in the design of molecular beacons. These tailored modifications expand the utility of oligos in complex experimental setups.
Quality Control and Purification
Ensuring the purity and accuracy of synthesized oligonucleotides is paramount. Impurities or sequence errors can lead to experimental inconsistencies. Standard purification methods include High-Performance Liquid Chromatography (HPLC) and Polyacrylamide Gel Electrophoresis (PAGE), both effective in removing truncated sequences and other contaminants. Advanced quality control measures, such as mass spectrometry, are also employed to verify molecular weight and sequence integrity.
Global Service Providers
Several organizations worldwide specialize in custom oligonucleotide synthesis, offering a range of services to meet diverse research needs. For instance, GenScript provides synthesis of DNA oligos up to 200 nucleotides in length, with over 200 available modifications. Similarly, Thermo Fisher Scientific offers a variety of custom DNA oligos, including primers and probes, with options for different scales and purifications. These providers utilize state-of-the-art technologies to deliver high-quality, customizable oligonucleotides to the global scientific community.