Biotin-dT - (10-1038)
5'-Dimethoxytrityloxy-5-[N-((4-t-butylbenzoyl)-biotinyl)-aminohexyl)-3-acrylimido]-2'-deoxyUridine-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
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10-1038
5'-Dimethoxytrityloxy-5-[N-((4-t-butylbenzoyl)-biotinyl)-aminohexyl)-3-acrylimido]-2'-deoxyUridine-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
Description
Biotin-dT can replace dT residues within the oligonucleotide sequence. 5'-Biotin phosphoramidite can be added ONLY ONCE to the 5'-terminus of an oligonucleotide. However, the DMT group on the biotin can be used in RP cartridge and HPLC purification techniques. PC Biotin is a photocleavable 5'-biotin phosphoramidite. BiotinTEG CPG and Protected BiotinLC Serinol CPG are designed for the direct synthesis of oligonucleotides containing biotin at the 3' terminus.
Desthiobiotin is a biotin analogue that exhibits lower binding to biotin-binding proteins such as streptavidin. This biotin analogue is lacking the sulfur group from the molecule and has a dissociation constant (Kd) several orders of magnitude less than biotin/streptavidin. As a result, biomolecules containing desthiobiotin are dissociated from streptavidin simply in the presence of buffered solutions of biotin. We offer desthiobiotinTEG phosphoramidite and the corresponding CPG.
Details
Usage
- Coupling: 15 minute coupling recommended
- Deprotection: No changes needed from standard method recommended by synthesizer manufacturer.
| Specifications | |
|---|---|
| Diluent | Anhydrous Acetonitrile |
| Recommended Storage | Refrigerated storage, maximum of 2-8°C, dry |
| Stability In Solution | 3-5 days |
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 | ||||
| 10-1038-02 | 0.25 g | .25grams | 1.94 | 51.33 | 30.8 | 19.25 | 14 | 10.27 | 2.57 |
| 10-1038-90 | 100 µmol | .129grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
| 10-1038-95 | 50 µmol | .064grams | 0.5 | 3.33 | 2 | 1.25 | 0.91 | 0.67 | 0.17 |
Expedite
| Catalog # | Pack Size | Grams/Pack | Dilution (mL) | Approximate Number of Additions | ||||
|---|---|---|---|---|---|---|---|---|
| Molarity | 50nm | 0.2μm | 1μm | 15μm | ||||
| 10-1038-02 | 0.25 g | .25grams | 2.9 | 0.07 | 51.6 | 32.25 | 23.45 | 3.23 |
| 10-1038-90 | 100 µmol | .129grams | 1.5 | 0.07 | 23.6 | 14.75 | 10.73 | 1.48 |
| 10-1038-95 | 50 µmol | .064grams | 0.75 | 0.07 | 8.6 | 5.38 | 3.91 | 0.54 |
Technical documents
Certificate of analysis
CoA search tool
Related products
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5'-Biotin Phosphoramidite
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DesthiobiotinTEG-CPG
Cat. No.: 20-2952
3'-BiotinTEG CPG
Cat. No.: 20-2955
3'-Protected BiotinLC Serinol CPG
Cat. No.: 20-2995
3'-BiotinTEG PS
Cat. No.: 26-2955
References
Products FAQS
1. Biotin dT will not affect the ability of kinase to phosphorylate at the 5'-terminus.2. The presence of Biotin-dT within an oligonucleotide does not affect hybridization relative to the same oligo containing dT(1). The biotin resides in the major groove of the formed double-stranded DNA and is readily available for binding to avidin or streptavidin.||REFERENCE(S): (1) J. Telser, K.A. Cruickshank, L.E. Morrison, and T.L. Netzel, J. Am. Chem. Soc., 1989, 111, 6966-6976. , ||
Biotin is transparent at 260nm. The UV detector in the HPLC trace of biotin phosphoramidites is monitoring the absorption of the DMT group on the spacer arm.||
No. However, this can be produced on the synthesizer by adding to the 5'- terminus first 5'-thiol-modifier C6 S-S (10-1936) followed by BioTEG phosphoramidite (10-1955). This should generate a biotinylated primer with a long spacer arm containing the disulfide linkage which can be cleaved later with DTT.||
A colorimetric assay for biotin can be quite effective. The color results from the reaction of biotin with p-dimethylaminocinnamaldehyde in the presence of sulfuric acid.1. Spot 0.2 A260 units of biotinylated oligonucleotide on a silica gel TLC plate.2. Dry the plate.]3. Spray with a solution of 2% p-dimethylaminocinnamaldehyde (Sigma), 2% conc. sulfuric acid in ethanol.4. Heat the plate and the presence of biotin will be indicated by the formation of a pink spot.Since the intensity of the biotin spot is quite low, it is prudent to compare with an unlabelled oligonucleotide similarly treated.
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