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5'-NPPOC Phosphoramidites
| Product Breadth of Proligo Reagents |
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| Product Breadth of Proligo Reagents |
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| Phosphoramidites with a NPPOC 5’-protection group can be used for the production of DNA microarrays using photolithographic in situ synthesis of oligonucleotides directly on the chip. This method offers the possibility to produce DNA chips of extremely high density and provides a flexible system. |
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View Structures:
NPPOC-dA(tac) Amidite
NPPOC-dC(ib) Amidite
NPPOC-dG(ipac) Amidite
NPPOC-dT Amidite |
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| Key Features of NPPOC Chemistry |
| Efficient photolytic deprotection (>99%) |
| Less side products during photolysis |
| Products available |
| Lot-to-lot consistent high purity and performance |
| Manufactured under a certified ISO 9001 quality system |
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| Analytical Specifications Test |
A |
C |
G |
T |
| HPLC Purity |
≤96.0% |
≤96.0% |
≤96.0% |
≤96.0% |
| 31P-NMR |
≤96.0% |
≤96.0% |
≤96.0% |
≤96.0% |
| Phosphoramidites |
| Product Information |
| Product No. |
Description |
Unit |
| A112N01-01 |
NPPOC-dA(tac) Amidite |
1 x 1g |
| C114N01-01 |
NPPOC-dC(ib) Amidite |
1 x 1g |
| G114N01-01 |
NPPOC-dG(ipac) Amidite |
1 x 1g |
| T111N01-01 |
NPPOC-dT Amidite |
1 x 1g |
User Instructions
1. Liquid Reagents
Deblock - The NPPOC-protective group is removed with light while the support-anchored oligonucleotide is immersed in a liquid reagent; various formulations are applicable, our recommended formulations are 1% NMI in DMSO (v/v) or 1% NMI in ACN (v/v) Activator - Any commercial activator that works for DNA-amidites is applicable; 0.25M DCI in ACN is perfect.
Oxidizer - Although any commercial oxidizer is applicable some customers use flow cells that are incompatible with THF and therefore prefer to use an oxidizer wherein the solvent THF is replaced by ACN.
Cap A - Fast Cap A is recommended (TAC-anhydride solution); some customers use flow cells that are incompatible with THF and therefore prefer to use a Cap A reagent wherein the solvent THF is replaced by ACN; some customers do not employ capping steps with NPPOC-chemistry.
Cap B - Cap B solutions that are compatible with Fast Cap A; some customers use flow cells that are incompatible with THF and therefore prefer to use a Cap B reagent wherein the solvent THF is replaced by ACN.
2. Coupling
NPPOC-amidites are dissolved in ACN at the concentration recommended for standard DNA amidites by the manufacturer of the DNA/RNA synthesizer. The coupling times of NPPOC-Amidites match the coupling times of standard DNA amidites (30 to 60 sec., longer coupling times are applicable, but not necessary).
3. Deprotection of NPPOC-groups
Light with a wavelength between 350 and 390 nm is applicable. The deprotection time depends on the light intensity. 90 sec. illumination time is sufficient at 100mW/cm2 on the surface of the synthesis support (365 nm Hg/Xe high pressure lamp).
4. Deprotection of base protective groups
Any fast deprotection protocol (e.g. conc. ammonia/room temp./2 hours or AMA/room temp./30 min.) is applicable. However, such treatment will partly remove the oligonucleotides from silica surfaces and therefore a milder treatment with a mixture of ethylendiamine and ethanol 50/50, v/v, for 2 hours at room temperature is used as an industry standard.
SAFC and SAFC Proligo Reagents are registered trademarks of Sigma-Aldrich Biotechnology, LP.
©2009 Sigma-Aldrich Biotechnology LP. All rights reserved. Reproduction forbidden without permission.
Other trademarks are the property of their respective owners.
To order or speak with a Customer Service Representative, contact your local SAFC office.
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