3'-Fluorescein-dT CPG

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20-2056
5'-Dimethoxytrityloxy-5-[N-((3',6'-dipivaloylfluoresceinyl)-aminohexyl)-3-acrylimido]-2'-deoxyUridine-3'-succinoyl-long chain alkylamino-CPG

Product Specifications

F.W.:
815.71
Catalog Number: Select Pack Size and Format to Reveal

Description

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 (TET)-, hexachloro (HEX) - and dichloro-dimethoxy (JOE)- 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.

Details

Usage

  • Coupling: This support should be used in a manner identical to normal protected nucleoside support since it contains the DMT group.
  • Deprotection: Use ammonium hydroxide and deprotect as required by nucleobases. When using AMA, a small amount of a non-fluorescent impurity will be formed. To eliminate this impurity, first deprotect with ammonium hydroxide for 30 minutes at room temperature, add an equal volume of 40% methylamine and then complete the deprotection as required by the nucleobases - e.g. 10 minutes at 65°C or 2 hours at room temperature for standard bases.
Specifications
Storage Freezer storage, -10 to -30°C, dry



Product FAQs

  • What are the relative extinction coefficients of 5'-Fluorescein, Hex and Tet etc.. at 260 nm and their Lambda max?
  • What are the relative extinction coefficients of various dyes?
  • Does AMA or methylamine cause any degradation to fluorescein or fluorescein-type dyes such as FAM or FITC?
  • Why does MALDI analysis of my oligos that contain one or more Fluorescein-dTs give an incorrect mass even though they give only a single, fluorescent band on a PAGE gel?