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Oligo Synthesis

Oligo Synthesis : Supports, CPGs

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3'-Fluorescein CPG

3'-Fluorescein CPG

Glen Research

Catalogue No.DescriptionPack SizePriceQty
20-2963-013'-Fluorescein CPG 0.1g £109.00 Quantity Add to Order
20-2963-103'-Fluorescein CPG 1.0g £905.00 Quantity Add to Order
20-2963-133'-Fluorescein CPG 1x10µm £273.00 Quantity Add to Order
20-2963-143'-Fluorescein CPG 1x15µm £409.00 Quantity Add to Order
20-2963-413'-Fluorescein CPG 4x1.0µm £182.00 Quantity Add to Order
20-2963-423'-Fluorescein CPG 4x0.2µm £109.00 Quantity Add to Order

Description

3'-Fluorescein CPG

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.

Structure

Catalog Number: 20-2963-xx

Description: 3'-Fluorescein CPG

1-Dimethoxytrityloxy-2-(N-thiourea-(di-O-pivaloyl-fluorescein)-
4-aminobutyl)-propyl-3-O-succinoyl-long chain alkylamino-CPG


F.W.: 598.56

Diluent: Not Applicable
Coupling: This support should be used in a manner identical to normal protected nucleoside support since it contains the DMT group.
Deprotection: Cleavage of the oligonucleotide from this support requires 2 hours at room temperature with ammonium hydroxide. Complete the deprotection using the protocol required by the nucleobases.
Storage: Freezer storage, -10 to -30°C, dry
Stability in Solution: Not Applicable

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Protocols

MSDS

 

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Notes

 3'-Fluorescein CPG

1-Dimethoxytrityloxy-2-(N-thiourea-(di-O-pivaloyl-fluorescein)-
4-aminobutyl)-propyl-3-O-succinoyl-long chain alkylamino-CPG
    F.W.: 598.56

Diluent: Not Applicable
Coupling: This support should be used in a manner identical to normal protected nucleoside support since it contains the DMT group.
Deprotection: Cleavage of the oligonucleotide from this support requires 2 hours at room temperature with ammonium hydroxide. Complete the deprotection using the protocol required by the nucleobases.
Storage: Freezer storage, -10 to -30°C, dry
Stability in Solution: Not Applicable
Material Safety Data Sheet

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.


If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications & Benefits

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.

relative extinction coefficients of various dyes

relative extinction coefficients of 5'-Fluorescein, Hex and Tet etc.. at 260 nm and their Lambda max

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Related products

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200