Custom Oligo Phosphorylation Services
In the development of life, nucleic acids (DNA and RNA) play a decisive role in biological macromolecules. It has been shown that nucleosides have strong anticancer and antiviral activities, and some nucleosides with modified glycosyl and base groups exhibit better biological activities. Oligonucleotides (Oligos) can be modified in various ways to utilize their active groups or to produce nucleoside analogs. One of the important modifications is phosphorylation.
Creative Biolabs can provide high-quality oligo modification services using our advanced synthesizer. We can achieve phosphorylation by adding phosphate groups to either 5'end or 3'end of an oligo.
5'-phosphorylation
If oligonucleotides are used as substrates for DNA ligases, 5'-phosphorylation is required. Oligonucleotides containing 5'-phosphate groups have multiple uses and are widely used in cloning, gene construction and ligation to ligate one oligonucleotide to another, for example as linkers and adapters. This is still the most common method of gene synthesis. This phosphate can be added enzymatically with a polynucleotide kinase or chemically added during oligonucleotide synthesis. As a chemical alternative to kinases, sulfoethyl phosphoramidites have been increasingly used in recent years. Because the chemical reagents are easy to synthesize, the yield of 5'-phosphate is usually much higher than that of kinases.
3'-phosphorylation
3'-phosphorylation is an efficient and commonly used for PCR blocking, which inhibits the degradation of certain 3'-exonucleases and can be used to block DNA polymerase activity. T4 polynucleotide kinase is widely used for 5'-phosphorylation of DNA and RNA oligonucleotide ends, but no natural protease is capable of 3'-phosphorylation. 3'- and 5'-phosphorylated oligonucleotides are used for further chemical modifications or as substrates for DNA repair studies. Chemical phosphorylation allows for bulk labeling and easier replication than enzymatic procedures.
Our Services
- we provide 5- and 3-phosphorylated oligos from 100 nmol up to 10 μmol.
- We can produce hundreds of customized nucleosides, including modified nucleosides and nucleoside polyphosphates, according to our customers' specifications.
- Analyze intermediates and final products with RP- and AX-HPLC, 31 p and 1 HNMR, mass spectrometry and LC-MS to ensure high purity and high quality.
- Phosphorylation requires additional processing to ensure high quality and high yield. For long oligonucleotides, PAGE purification is recommended and extended HPLC can also be used.
Delivery:
- Tube or plate formats
- Oligos are delivered in lyophilized format
Advantages:
- Rich Experience
- High Quality - Strict quality control system with low error rates
- Fast delivery: deliver within 2 weeks
- Reports: we provide reports containing O.D. values, nmol, M.V. and TM of oligonucleotides.
- Competitive pricing
- Advanced mass spectrometry QC available for oligo modification
Creative Biolabs can supply professional customized services on chemical modifications of nucleic acid drugs and support your research and technology applications. Please contact us to learn more.
Reference
- An antisense oligonucleotide primer. Hogrefe RI; Antisense and Nucleic Acid Drug Development (1999), 9(4): 351-357.
*For Research Use Only. Not for use in diagnostic procedures.