5'-Dichloro-dimethoxy-Fluorescein Phosphoramidite (Joe) |
Catalog Number: 10-5904-xx
Description: 5'-Dichloro-dimethoxy-Fluorescein Phosphoramidite (Joe)
4-[4',5'-Dichloro-2',7'-dimethoxy-3',6'-dicyclohexanecarboxylfluorescein-6-yl- carboxamido]-cyclohexyl-1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite |
Formula: C52H62N3O12Cl2P |
M.W.: 1022.94 |
F.W.: 665.39 |
Diluent: Anhydrous Acetonitrile |
Coupling: 6 minute coupling time recommended |
Deprotection: Use Ammonium Hydroxide and deprotect as required by nucleobases. |
Storage: Freezer storage, -10 to -30°C, dry |
Stability in Solution: 1-2 days |
FLUORESCEIN LABELLING
5’-Fluorescein phosphoramidite contains no 4,4’-dimethoxytrityl (DMT) group and can be added only once at the 5’-terminus, thereby terminating synthesis. This product is prepared using the 6-carboxyfluorescein derivative. The tetrachloro-, hexachloro-and dichloro-dimethoxy-fluorescein phosphoramidites are designed to take advantage of the multicolor detection capability of modern DNA sequencers and genetic analyzers. Fluorescein phosphoramidite is designed to produce the same fluorescein-type structure as had been previously prepared using fluorescein isothiocyanate (FITC). Our fluorescein phosphoramidite also contains a DMT group to allow quantification of coupling. The analogous structure, 6-Fluorescein Phosphoramidite, prepared using 6-FAM, is also available, along with 6-Fluorescein Serinol Phosphoramidite. Fluorescein-dT can be inserted into the desired sequence as a replacement for a dT residue.
We offer five fluorescein supports. Fluorescein CPG has traditionally been used to add the fluorescein label at the 3’-terminus. The analogous structure, 3’-(6-Fluorescein) CPG, prepared using 6-FAM, is now also available, along with 6-Fluorescein Serinol CPG. We also offer 3’-(6-FAM) CPG and Fluorescein-dT CPG, both derivatives of 6-carboxyfluorescein (6-FAM). Both are single isomers and use an amide linkage which is stable during cleavage and deprotection and does not allow isomer formation. 3’-(6-FAM) CPG allows effective blockage of the 3’-terminus from polymerase extension as well as exonuclease digestion. Fluorescein-dT CPG allows both of these enzymatic activities to proceed. Normal cleavage and deprotection with ammonium hydroxide readily generates the fluorescein labelled oligos.
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