Oligo Synthesis : CEPs
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5"-Biotin Phosphoramidite
5"-Biotin Phosphoramidite
Glen Research
| Catalogue No. | Description | Pack Size | Price | Qty |
|
|---|---|---|---|---|---|
| 10-5950-02 | 5"-Biotin Phosphoramidite | 0.25g | £260.00£247.00Offer until : 31-Mar-2021Offer Code : GLEN5View Offer | Quantity | Add to Order |
| 10-5950-90 | 5"-Biotin Phosphoramidite | 100µmoles | £120.00£114.00Offer until : 31-Mar-2021Offer Code : GLEN5View Offer | Quantity | Add to Order |
| 10-5950-95 | 5"-Biotin Phosphoramidite | 50µmoles | £80.00£76.00Offer until : 31-Mar-2021Offer Code : GLEN5View Offer | Quantity | Add to Order |
Related products
5"-Biotin Phosphoramidite
5"-Biotin Phosphoramidite
Glen Research
| 5"-Biotin Phosphoramidite |
Catalog Number: 10-5950-xx
Description: 5"-Biotin Phosphoramidite
| [1-N-(4,4"-Dimethoxytrityl)-biotinyl-6-aminohexyl]-2-cyanoethyl-(N,N-diisopropyl)-phosphoramidite | ||
| Formula: C46H64N5O6PS | M.W.: 846.08 | F.W.: 405.45 |
Diluent: Anhydrous Acetonitrile |
| 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. |
| Storage: Freezer storage, -10 to -30°C, dry |
Stability in Solution: 1-2 days Biotin-dT can replace dT residues within the oligonucleotidesequence. 5’-Biotin phosphoramidite can be added ONLY ONCE to the 5’-terminus ofan oligonucleotide. However, the DMT group on the biotin can be used in RPcartridge and HPLC purification techniques. PC Biotin is a photocleavable5’-biotin phosphoramidite. BiotinTEG CPG and Protected BiotinLC Serinol CPG aredesigned for the direct synthesis of oligonucleotides containing biotin at the3’ terminus. Desthiobiotin is a biotin analogue that exhibits lower binding tobiotin-binding proteins such as streptavidin. This biotin analogue is lackingthe sulfur group from the molecule and has a dissociation constant (Kd) severalorders of magnitude less than biotin/streptavidin. As a result, biomoleculescontaining desthiobiotin are dissociated from streptavidin simply in thepresence of buffered solutions of biotin. We offer desthiobiotinTEGphosphoramidite and the corresponding CPG. ABI-style vials and columns are supplied unless otherwise requested (see notebox). |
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5"-Biotin Phosphoramidite
5"-Biotin Phosphoramidite
Glen Research
MSDS
Glen Report 8.2: 5" LABELLING - FLUORESCEIN AND CYANINE DYES, BIOTIN
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5"-Biotin Phosphoramidite
5"-Biotin Phosphoramidite
Glen Research
Frequently Asked Technical QuestionQUESTION: 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: What absorbance does biotin have at 260nm?The HPLC trace shows absorbance at 254nm? RESPONSE: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. QUESTION: Do you have a biotin phosphoramidite containing a disulfide linker which can be cleaved later with DTT to release the DNA from a streptavidin support? RESPONSE: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 spacerarm containing the disulfide linkage which can be cleaved later with DTT. QUESTION: How can I tell if the biotinylated oligonucleotide I have made really does contain biotin? RESPONSE: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. 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 DATAThe 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.
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If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200
