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Catalog Number: 10-5901-xx
Description: 5'-Fluorescein Phosphoramidite
| 6-(3',6'-dipivaloylfluoresceinyl-6-carboxamido)-hexyl- 1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite | ||
| Formula: C46H58N3O10P | M.W.: 843.95 | F.W.: 537.46 |
Diluent: Anhydrous Acetonitrile |
| Coupling: 3 minute coupling time recommended |
| Deprotection: No changes needed from standard method recommended by synthesizer manufacturer. Technical Bulletin |
| Storage: Freezer storage, -10 to -30°C, dry |
| Stability in Solution: 1-2 days, <90% efficient after 4 days |
| Please Note: This product or portions thereof is manufactured under license from GE Healthcare Bio-Sciences Corp. under US Patent Numbers 5,371,241, 5,721,355, and/or 5,583,236. |
| Catalog Information | Material Safety Data Sheet |
Literature Highlights
Frequently Asked Technical Question
QUESTION: What are the relative extinction coefficients of 5'-Fluorescein, Hex and Tet etc.. at 260 nm and their Lambda max?
RESPONSE:Please see http://www.glenresearch.com/Technical/Extinctions.html
REFERENCE(S):
Oligonucleotide Properties Calculator; http://www.basic.northwestern.edu/biotools/oligocalc.html
QUESTION: What are the relative extinction coefficients of various dyes?
RESPONSE:Please see http://www.glenres.com/Technical/Extinctions.html#dyes
QUESTION: Does AMA or methylamine cause any degradation to fluorescein or fluorescein-type dyes such as FAM or FITC?
RESPONSE:Response: While AMA (Ammonium hydroxide/40% Methylamine 1:1 v/v) is considered compatible with fluorescein, the use of methylamine when deprotecting a Fluorescein-labeled oligo does lead to a small amount of degradation, which is characterized by a the appearance of a late-eluting peak by RP HPLC that shows no visible fluorescein absorbance. With standard deprotection conditions (AMA 10 minutes at 65 C) the amount of degradation is approximately 5%. The impurity is not detected with AMA at RT for 2 hours.
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. | Pack Size | Grams/ Pack | 0.1M Dil. (mL) | LV40 | LV200 | 40nm | 0.2µm | 1µm | 10µm |
| Approximate Number of Additions | |||||||||
| 10-5901-02 | 0.25grams | .25grams | 2.96 | 85.33 | 51.2 | 32 | 23.27 | 17.07 | 4.27 |
| 10-5901-90 | 100µmoles | .084grams | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
| 10-5901-95 | 50µmoles | .042grams | .5 | 3.33 | 2 | 1.25 | .91 | .67 | .17 |
| Expedite | |||||||||
| Cat.No. | Pack Size | Grams/ Pack | Dilution (mL) | Molarity | 50nm | 0.2µm | 1µm | 15µm | |
| Approximate Number of Additions | |||||||||
| 10-5901-02 | 0.25grams | .25grams | 4.42 | .07 | 82 | 51.25 | 37.27 | 5.13 | |
| 10-5901-90 | 100µmoles | .084grams | 1.5 | .07 | 23.6 | 14.75 | 10.73 | 1.48 | |
| 10-5901-95 | 50µmoles | .042grams | .75 | .07 | 8.6 | 5.38 | 3.91 | .54 | |
| Beckman | |||||||||
| Cat.No. | Pack Size | Grams/ Pack | Dilution (mL) | Molarity | 30nm | 200nm | 1000nm | ||
| Approximate Number of Additions | |||||||||
| 10-5901-02 | 0.25grams | .25grams | 4.42 | .07 | 83.6 | 52.25 | 38 | ||
| 10-5901-90 | 100µmoles | .084grams | 1.5 | .07 | 25.2 | 15.75 | 11.45 | ||
| 10-5901-95 | 50µmoles | .042grams | .75 | .07 | 10.2 | 6.38 | 4.64 | ||
08/15/2012 | http://www.glenres.com/ProductFiles/10-5901.html