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Catalog Number: 10-1985-xx
Description: DNP-TEG Phosphoramidite
| 1-Dimethoxytrityloxy-3-O-[N-(2,4-dinitrophenyl)-3-N-aminopropyl-(triethyleneglycol)]- glyceryl-2-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite | ||
| Formula: C48H64N5O13P | M.W.: 950.00 | F.W.: 509.41 |
Diluent: Anhydrous Acetonitrile Add fresh diluent to product vial to recommended concentration and swirl vial occasionally over several minutes until product is completely dissolved. (Some oils may require between 5 and 10 minutes.) Use care to maintain anhydrous conditions. In case of transfer to alternate vial type, ensure recipient vial has been pre-dried. For more information, see http://www.glenres.com/Technical/TB_ABITransfer.pdf. |
| Coupling: 12-15 minute coupling time. To maintain label yield, carry out the synthesis DMT-on. |
| Deprotection: No changes needed from standard method recommended by synthesizer manufacturer. To maintain label yield, remove the 5'-DMT after ammonium hydroxide deprotection. |
| Storage: Freezer storage, -10 to -30°C, dry |
| Stability in Solution: 2-3 days |
| Catalog Information | Material Safety Data Sheet |
Literature Highlights
| Glen Report 6.1: NEW LABELLING REAGENTS |
Frequently Asked Technical Question
QUESTION: Can oligonucleotides modified at the 5'-terminus with, for example, biotin be phosphorylated with kinase?
RESPONSE:Modification reagents based on a 1,2-diol, e.g., BioTEG, DNP phosphoramidites, or a 1,3-diol, e.g., fluorescein, biotin phosphoramidites, can be added several times at the 5'-terminus since they contain an alcohol group capable of further addition with phosphoramidites. Can this alcohol also be as a substrate for T4 polynucleotide kinase for 32P labelling of these modified oligonucleotides? Surprisingly, the answer is yes. Teoule and coworkers have shown(1) that oligos labelled at the 5'-terminus are substrates for kinase. Interestingly, the oligos modified with reagents based on 1,2-diols are labelled to 50%, indicating that only one diastereomer is labelled, while those modified with 1,3-diol reagents are labelled to 100%.
REFERENCE(S):
(1) M.L. Fontanel, H. Bazin, and R. Teoule, Analytical Biochemistry, 1993, 214, 338-340.
QUESTION: Can oligonucleotides modified at the 5'-terminus with, for example, biotin be phosphorylated with kinase?
RESPONSE:Modification reagents based on a 1,2-diol, e.g., BioTEG, DNP phosphoramidites, or a 1,3-diol, e.g., fluorescein, biotin phosphoramidites, can be added several times at the 5'-terminus since they contain an alcohol group capable of further addition with phosphoramidites. Can this alcohol also be as a substrate for T4 polynucleotide kinase for 32P labelling of these modified oligonucleotides? Surprisingly, the answer is yes. Teoule and coworkers have shown(1) that oligos labelled at the 5'-terminus are substrates for kinase. Interestingly, the oligos modified with reagents based on 1,2-diols are labelled to 50%, indicating that only one diastereomer is labelled, while those modified with 1,3-diol reagents are labelled to 100%.
REFERENCE(S):
(1) M.L. Fontanel, H. Bazin, and R. Teoule, Analytical Biochemistry, 1993, 214, 338-340.
DILUTION/COUPLING DATA
The table below shows pack size data and, for solutions, dilutions and approximate couplings based on normal priming procedures. Please link for more detailed usage information with the various synthesizers.
| ABI 392/394 | |||||||||
| Cat.No. | Pack Size | Grams/ Pack | 0.1M Dil. (mL) | LV40 | LV200 | 40nm | 0.2µm | 1µm | 10µm |
| Approximate Number of Additions | |||||||||
| 10-1985-02 | 0.25grams | .25grams | 2.63 | 74.33 | 44.6 | 27.88 | 20.27 | 14.87 | 3.72 |
| 10-1985-90 | 100µmoles | .095grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
| 10-1985-95 | 50µmoles | .048grams | .5 | 3.33 | 2 | 1.25 | .91 | .67 | .17 |
| Expedite | |||||||||
| Cat.No. | Pack Size | Grams/ Pack | Dilution (mL) | Molarity | 50nm | 0.2µm | 1µm | 15µm | |
| Approximate Number of Additions | |||||||||
| 10-1985-02 | 0.25grams | .25grams | 3.93 | .07 | 72.2 | 45.13 | 32.82 | 4.51 | |
| 10-1985-90 | 100µmoles | .095grams | 1.5 | .07 | 23.6 | 14.75 | 10.73 | 1.48 | |
| 10-1985-95 | 50µmoles | .048grams | .75 | .07 | 8.6 | 5.38 | 3.91 | .54 | |
| Beckman | |||||||||
| Cat.No. | Pack Size | Grams/ Pack | Dilution (mL) | Molarity | 30nm | 200nm | 1000nm | ||
| Approximate Number of Additions | |||||||||
| 10-1985-02 | 0.25grams | .25grams | 3.93 | .07 | 73.8 | 46.13 | 33.55 | ||
| 10-1985-90 | 100µmoles | .095grams | 1.5 | .07 | 25.2 | 15.75 | 11.45 | ||
| 10-1985-95 | 50µmoles | .048grams | .75 | .07 | 10.2 | 6.38 | 4.64 | ||
08/15/2012 | http://www.glenres.com/ProductFiles/10-1985.html