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Oligo Synthesis

Oligo Synthesis : CEPs

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5'-Dichloro-dimethoxy-Fluorescein Phosphoramidite (Joe)

5'-Dichloro-dimethoxy-Fluorescein Phosphoramidite (Joe)

Glen Research

Description

5'-Dichloro-dimethoxy-Fluorescein Phosphoramidite (Joe)

Structure

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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Protocols

MSDS

Glen Report 20.2

Glen Report 21.2

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Applications & Benefits

DILUTION/COUPLING DATA

The table below shows pack size data and, for solutions, dilutions and approximate couplings based on normal priming procedures.

ABI 392/394
Cat.No. Pack
Size
Grams/
Pack
0.1M Dil.
(mL)
LV40 LV200 40nm 0.2µm 1µm 10µm
Approximate Number of Additions
10-5904-02 0.25grams .25grams 2.44 68 40.8 25.5 18.55 13.6 3.4
10-5904-90 100µmoles .102grams 1 20 12 7.5 5.45 4 1
10-5904-95 50µmoles .051grams .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
10-5904-02 0.25grams .25grams 3.65 .07 66.6 41.63 30.27 4.16  
10-5904-90 100µmoles .102grams 1.5 .07 23.6 14.75 10.73 1.48  
10-5904-95 50µmoles .051grams .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
10-5904-02 0.25grams .25grams 3.65 .07 68.2 42.63 31    
10-5904-90 100µmoles .102grams 1.5 .07 25.2 15.75 11.45    
10-5904-95 50µmoles .051grams .75 .07 10.2 6.38 4.64

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Related products

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