5'-Biotin Phosphoramidite - (10-5950)
[1-N-(4,4'-Dimethoxytrityl)-biotinyl-6-aminohexyl]-2-cyanoethyl-(N,N-diisopropyl)-phosphoramidite
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10-5950
[1-N-(4,4'-Dimethoxytrityl)-biotinyl-6-aminohexyl]-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: 2 minute coupling time recommended
- Deprotection: 5'-Biotin is slow to detritylate. If the final DMT-group is to be removed on the synthesizer, we recommend a second deblocking step. If the final DMT-group is left on to aid in cartridge purification, we recommend that the oligo is left in contact with the TFA solution for 10 minutes.
| Specifications | |
|---|---|
| Diluent | Anhydrous Acetonitrile |
| Recommended Storage | Freezer storage, -10 to -30°C, dry |
| Stability In Solution | 1-2 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 | ||||
| 10-5950-02 | 0.25 g | .25grams | 2.95 | 85 | 51 | 31.88 | 23.18 | 17 | 4.25 |
| 10-5950-90 | 100 µmol | .085grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
| 10-5950-95 | 50 µmol | .042grams | 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 | ||||
| 10-5950-02 | 0.25 g | .25grams | 4.41 | 0.07 | 81.8 | 51.13 | 37.18 | 5.11 |
| 10-5950-90 | 100 µmol | .085grams | 1.5 | 0.07 | 23.6 | 14.75 | 10.73 | 1.48 |
| 10-5950-95 | 50 µmol | .042grams | 0.75 | 0.07 | 8.6 | 5.38 | 3.91 | 0.54 |
Technical documents
Certificate of analysis
CoA search tool
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Products FAQS
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.
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 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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