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Oligo Synthesis

Oligo Synthesis : CEPs

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DNP-TEG Phosphoramidite

DNP-TEG Phosphoramidite

Glen Research

Catalogue No.DescriptionPack SizePriceQty
10-1985-02DNP-TEG Phosphoramidite 0.25g £614.00 Quantity Add to Order
10-1985-90DNP-TEG Phosphoramidite 100µmoles £268.00 Quantity Add to Order
10-1985-95DNP-TEG Phosphoramidite 50µmoles £150.00 Quantity Add to Order

Description

DNP-TEG Phosphoramidite

Structure

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

DNP LABELLING

An analytical test based on detection of 2,4-dinitrophenyl (DNP) labelled oligonucleotides with anti-DNP antibodies has been proposed. We have chosen the branched triethylene glycol (TEG) spacer in our version of DNP phosphoramidite since it can be added once or several times to the 3’ or 5’ terminus.

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Protocols

Material Safety Data Sheet

Glen Report 6.1: NEW LABELLING REAGENTS

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Notes

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.

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Applications & Benefits

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

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