Fluorescein-dT phosphoramidite

Product Introduction

This phosphoramidite allows fluorescein (FAM) to be introduced into any position in the oligonucleotide sequence during synthesis by the phosphoramidite method (in the middle of the chain, at the 5’- and
3’-terminus). The reagent is a conjugate of deoxythymidine phosphoramidite
and 6-isomer of FAM. Modification is performed during oligonucleotide synthesis by substituting standard dT phosphoramidite with fluorescein-dT phosphoramidite. This modification does not affect exonuclease or polymerase activity.,For modification with fluorescein at the 5’-terminus use
FAM phosphoramidite, 6-isomer.,Coupling: 10 minutes,Deprotection: standard conditions using ammonium hydroxide;
deprotection time depends on oligonucleotide composition and nucleobase
protecting groups (deprotection for 17 h at 55 °С removes all protecting
groups from standard nucleobases). AMA (solution of 30% ammonium
hydroxide/40% aqueous methylamine 1:1 v/v) can be used with ~5% of
non-fluorescent side product forming. To avoid formation of the side product, start deprotection with ammonium hydroxide (30 min at room temperature), then add an equal volume of 40% aqueous methylamine and continue deprotection as required with AMA (e.g. 10 min at 65 °C).

Chemical Information
Purity	NMR 1H, NMR 31P, HPLC-MS
Solubility	Good in acetonitrile, DCM
Appearance	Off White powder

Product Specification
CF260	0.20
CF280	0.17
Fluorescence Quantum Yield	0.93
Excitation	490
Emission	513
Storage	Storage: 12 months after receival at -20 °C in the Dark. Transportation: at room temperature for up to 3 weeks. Avoid prolonged exposure to light. Desiccate.