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Chemical Phosphorylation
Oligonucleotides containing a 5'-phosphate group have various applications, and are often used as a means of ligating one oligo to another, e.g. as linkers and adapters, in cloning, gene construction, and ligation in general.
3'-Phosphorylations are used to block enzyme activity. For example, this is an efficient and commonly used PCR blocking technique.
5’-Phosphate Modifications
Phosphorylation of the 5'-terminus on oligonucleotides is routinely achieved, with higher yields than using kinase, using a Chemical Phosphorylation Reagent (also known as CPR or “Phosphate-ON”)1.
Chemical phosphorylation reagents for 5'-terminus modification
Product | Description and Advantages |
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Chemical Phosphorylating Reagent (CPR) NACE1-008 |
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5'-Phosphorylating Amidite II NACE1-006 |
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Chemical Phosphorylating Reagent (CPR II)2 NACE1-007 |
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Solid Chemical Phosphorylating Reagent (solid CPR)3 NACE1-011 |
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3' Phosphate Modification of an Oligonucleotide
3'-Phosphate CPGs allow direct preparation of oligonucleotides with a 3'-phosphate group.
Chemical phosphorylation reagents for 3'-terminus modification
Product | Pore Size Availability |
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3'-Phosphate CPG NACE1-002 |
500 Å, 1000 Å, 3000 Å |
3'-Phosphate CPG Column NACE1-003 |
1000 Å, 3000 Å (available in different column formats) |
DMT-Phosphate-Suc-CPG NACE1-009 |
500 Å, 1000 Å |
DMT-Phosphate-Suc-CPG Column Suc-CPG) NACE1-010 |
1000 Å, 1400 Å (available in different column formats) |
Our 3’-Phosphate CPGs are available in different pore sizes:
CPG Pore Size | Application |
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500 Å | Suitable for high yield applications such as therapeutic oligos (≤ 30mers) |
1000 Å | Suitable for highly modified oligonucleotides (> 20mers) |
3000 Å | Performs well for very long sequences (> 120mers) |
References
- A chemical 5'-phosphorylation of oligodeoxyribonucleotides that can be monitored by trityl cation release, T. Horn and M. Urdea, Tetrahedron Lett., 27, 4705-4708, 1986.
- A new approach for chemical phosphorylation of oligonucleotides at the 5'-terminus, A. Guzaev, H. Salo, A. Azhayev and H. Lonnberg, Tetrahedron, 51, 9375-9384, 1995.
- Chemical phosphorylation of oligonucleotides and reactants therefor, A. Guzaev, A. Azhayev and H. Lonnberg, US Patent No. 5959090, 1999.