5'-Dabsyl-dT-CE Phosphoramidite - (10-1532)
5'-Dabsyl-2'-deoxyThymidine,3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
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10-1532
5'-Dabsyl-2'-deoxyThymidine,3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
Description
QUAL probes1 consist of two oligonucleotides, the first containing a nucleophilic group at the 3'-terminus, while the second has an electrophilic group at the 5'-terminus. When the probe pair finds the target, the oligos line up with the 3'-terminus of the first directly adjacent to the 5'-terminus of the second. An autoligation reaction then takes place to combine the two oligos into a single probe. As usual, the 3' nucleophilic group is the 3-thiophosphate, easily prepared using 3'-phosphate CPG with a sulfurizing step in the first cycle. In this case, the electrophilic group is a 5'-dabsyl group, which is an excellent leaving group as well as a fine quencher of fluorescence. The second oligo, therefore, contains a fluorophore which is quenched by the dabsyl group. A popular choice for fluorophore is fluorescein-dT but it is easy to imagine that a variety of fluorophores could be attached to any of the commercially available amino-modified nucleoside phosphoramidites.
Details
Usage
- Coupling: Monomers that allow for UltraMILD deprotection must be used. (dA: 10-1601-xx, dC: 10-1015-xx, dG: 10-1621-xx, dT: 10-1030-xx ). To avoid any exchange of the iPr-Pac group on the dG|with acetyl, use the UltraMild Cap Mix A (40-4210-xx/ 40-4212-xx).
- Deprotection: UltraMILD 0.05M Potassium Carbonate in Methanol, 4 hours at Room Temperature.
Specifications | |
---|---|
Diluent | Anhydrous Acetonitrile |
Recommended Storage | Freezer storage, -10 to -30°C, dry |
Stability In Solution | 1-2days |
Dilution/Coupling Data
The table below show pack size data and, for solutions, dilution and approximate coupling based on normal priming procedures.
ABI 392/394
Catalog # | Pack Size | Grams/Pack | 0.1M Dil. (mL) | Approximate Number of Additions | |||||
---|---|---|---|---|---|---|---|---|---|
LV40 | LV200 | 40nm | 0.2μm | 1μm | 10μm | ||||
10-1532-02 | 0.25 g | .25grams | 3.43 | 101 | 60.6 | 37.88 | 27.55 | 20.2 | 5.05 |
10-1532-90 | 100 µmol | .073grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
Expedite
Catalog # | Pack Size | Grams/Pack | Dilution (mL) | Approximate Number of Additions | ||||
---|---|---|---|---|---|---|---|---|
Molarity | 50nm | 0.2μm | 1μm | 15μm | ||||
10-1532-02 | 0.25 g | .25grams | 5.11 | 0.07 | 95.8 | 59.88 | 43.55 | 5.99 |
10-1532-90 | 100 µmol | .073grams | 1.5 | 0.07 | 23.6 | 14.75 | 10.73 | 1.48 |
Technical documents
Certificate of analysis
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