Oligo Synthesis : CEPs
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BiotinTEG Phosphoramidite
BiotinTEG Phosphoramidite
Glen Research
| Catalogue No. | Description | Pack Size | Price | Qty |
|
|---|---|---|---|---|---|
| 10-1955-02 | BiotinTEG Phosphoramidite | 0.25g | £500.00£475.00Offer until : 31-Mar-2021Offer Code : GLEN5View Offer | Quantity | Add to Order |
| 10-1955-90 | BiotinTEG Phosphoramidite | 100µmoles | £236.00£224.20Offer until : 31-Mar-2021Offer Code : GLEN5View Offer | Quantity | Add to Order |
| 10-1955-95 | BiotinTEG Phosphoramidite | 50µmoles | £118.00£112.10Offer until : 31-Mar-2021Offer Code : GLEN5View Offer | Quantity | Add to Order |
Related products
BiotinTEG Phosphoramidite
BiotinTEG Phosphoramidite
Glen Research
| BiotinTEG Phosphoramidite |
Catalog Number: 10-1955-xx
Description: BiotinTEG Phosphoramidite
| 1-Dimethoxytrityloxy-3-O-(N-biotinyl-3-aminopropyl)-triethyleneglycolyl-glyceryl-2-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite | ||
| Formula: C52H76N5O11PS | M.W.: 1010.24 | F.W.: 569.61 |
Diluent: Anhydrous Acetonitrile |
| 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: Refrigerated storage, maximum of 2-8°C, dry |
| Stability in Solution: 2-3 days |
BIOTIN LABELLING
Glen Research biotin phosphoramidites for direct labelling of syntheticoligonucleotides exhibit the following features:
1. All are soluble in acetonitrile at concentrations useful for DNAsynthesis.
2. All include a DMT group for cartridge purifications which is essential forthe preparation of biotinylated PCR primers because of the potential for crosscontamination in HPLC purifications.
3. For the development of diagnostic probes, biotin phosphoramidite iscapable of branching to allow multiple biotins to be introduced at the 3’- or5’-terminus while biotin-dT can replace dT residues within the oligonucleotidesequence. BiotinTEG Phosphoramidite contains a 15 atom mixed polarity spacer armbased on a triethylene glycol. 5’-Biotin phosphoramidite can be added ONLY ONCEto the 5’-terminus of an oligonucleotide. However, the DMT group on the biotincan be used in RP cartridge and HPLC purification techniques.
4. Protected Biotin Serinol Phosphoramidite and CPG are protected with at-butylbenzoyl group on the biotin ring. This group is designed to stop anyphosphoramidite reactions at this active position in biotin. This protectionavoids branching when using nucleophilic activators like DCI. The protectinggroup is easily removed during oligonucleotide cleavage and deprotection. TheBiotinLC versions are similarly protected and should be useful for the synthesisof highly sensitive biotinylated probes.
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BiotinTEG Phosphoramidite
BiotinTEG Phosphoramidite
Glen Research
Material Safety Data Sheet
Glen Report 6.2: RESEARCH REVIEW - ANTISENSE RNA
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BiotinTEG Phosphoramidite
BiotinTEG Phosphoramidite
Glen Research
Literature Highlights
| Glen Report 6.2: RESEARCH REVIEW - ANTISENSE RNA |
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: 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.
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BiotinTEG Phosphoramidite
BiotinTEG Phosphoramidite
Glen Research
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. | PackSize | Grams/Pack | 0.1M Dil.(mL) | LV40 | LV200 | 40nm | 0.2µm | 1µm | 10µm |
| Approximate Number of Additions | |||||||||
| 10-1955-02 | 0.25grams | .25grams | 2.47 | 69 | 41.4 | 25.88 | 18.82 | 13.8 | 3.45 |
| 10-1955-90 | 100µmoles | .101grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
| 10-1955-95 | 50µmoles | .051grams | .5 | 3.33 | 2 | 1.25 | .91 | .67 | .17 |
| Expedite | |||||||||
| Cat.No. | PackSize | Grams/Pack | Dilution(mL) | Molarity | 50nm | 0.2µm | 1µm | 15µm | |
| Approximate Number of Additions | |||||||||
| 10-1955-02 | 0.25grams | .25grams | 3.69 | .07 | 67.4 | 42.13 | 30.64 | 4.21 | |
| 10-1955-90 | 100µmoles | .101grams | 1.5 | .07 | 23.6 | 14.75 | 10.73 | 1.48 | |
| 10-1955-95 | 50µmoles | .051grams | .75 | .07 | 8.6 | 5.38 | 3.91 | .54 | |
| Beckman | |||||||||
| Cat.No. | PackSize | Grams/Pack | Dilution(mL) | Molarity | 30nm | 200nm | 1000nm | ||
| Approximate Number of Additions | |||||||||
| 10-1955-02 | 0.25grams | .25grams | 3.69 | .07 | 69 | 43.13 | 31.36 | ||
| 10-1955-90 | 100µmoles | .101grams | 1.5 | .07 | 25.2 | 15.75 | 11.45 | ||
| 10-1955-95 | 50µmoles | .051grams | .75 | .07 | 10.2 | 6.38 | 4.64 | ||
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