Catalog Number: 26-2961-xx
Description: 3'-(6-FAM) PS
Diluent: Anhydrous Acetonitrile
|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: 2-3 days
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.
All Glen Research CPG supports use the standard long chain alkylamino (lcaa) linker but differ in the glass pore size, 500Å, 1000Å or 2000Å. The 500Å support is appropriate for shorter sequences, while the 1000Å supports perform better in the synthesis of longer (>30-mer) DNA sequences. The 2000Å support is best for very long (>150-mer) oligonucleotides. We have instituted an additional QC test for supports to show the length of oligo that can be prepared before a drop-off in coupling due to steric effects begins to occur. The drop-off point is recorded in the Certificate of Analysis. All Glen Research supports are fully end-capped to ensure that the CPG surface is totally inert, thereby avoiding the introduction of impurity sequences containing deletions at the 3’-terminus.
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