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5-I-dU-CE Phosphoramidite - (10-1091)

5'-Dimethoxytrityl-5-iodo-2'-deoxyUridine,3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
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10-1091
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Product Specifications

Formula:
C39H46IN4O8P
M.W.:
856.69
F.W.:
416.07
CAS Number:
178925-48-9
5'-Dimethoxytrityl-5-iodo-2'-deoxyUridine,3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
Description
Brominated and iodinated nucleosides are used in crystallography studies of oligonucleotide structure. They are also photolabile and are used for cross-linking studies to probe the structure of protein-DNA complexes. Antibodies exist to Br-dU and oligonucleotides containing Br-dU can be used as probes.
Details

Usage

  • Coupling: No changes needed from standard method recommended by synthesizer manufacturer.
  • Deprotection: Mild Ammonium Hydroxide for 24 hours at room temperature
Specifications
Diluent Anhydrous Acetonitrile
Recommended Storage Refrigerated storage, maximum of 2-8°C, dry
Stability In Solution 24 hours
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
10-1091-02 0.25 g .25grams 2.92 84 50.4 31.5 22.91 16.8 4.2
10-1091-90 100 µmol .086grams 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
10-1091-02 0.25 g .25grams 4.36 0.07 80.8 50.5 36.73 5.05
10-1091-90 100 µmol .086grams 1.5 0.07 23.6 14.75 10.73 1.48
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Products FAQS

Please see: Extinction Table

REFERENCE(S):B. Froehler, Gilead, M. Powell, R. Somers

The most widely used method seems to be using Br-dU and, more recently, I-dU (1). The substitution of photoreactive Br and I for the 5-Me group of thymidine is attractive since their radii are similar to that of the methyl group. These modifications do not significantly affect the binding of oligonucleotides with proteins. Br-dU is irradiated at 308nm and crosslinking is typically not greater than 40%. Crosslinking of I-dU at 308nm is higher but is optimal at 325nm. Laser excitation is preferred.After the synthesis of oligonucleotides containing Br-dU or I-dU, care must be taken to avoid loss of the halogens. Even though both modifications are quite stable to ammonium hydroxide, it is sensible to play safe and carry out the deprotection at room temperature for 24 hours. Even better, use the UltraMild monomers and deprotect with potassium carbonate in methanol at room temeperature. The oligonucleotide products should be protected from light - plastic tubes and amber vials are safe. Gel electrophoresis should be carried out protected from light.||REFERENCE(S):(1) M.C. Willis, B.J. Hicke, O.C. Uhlenbeck, T.R. Cech and T.H. Koch, Science, 1993, 262, 1255.||