6-Fluorescein phosphoramidite
There are several ways of labeling an oligonucleotide with fluorescein. The choice of label is diversified further, depending on the spectral requirements. 5'-fluorescein-CE phosphoramidite, derived from the single isomer 6-carboxyfluorescein, 5'-hexachlorofluorescein-CE phosphoramidite (HEX) and 5'-tetrachlorofluorescein-CE phosphoramidite (TET) can all be used to efficiently label an oligonucleotide at the 5'-end. Labeling the 3'-end of an oligo with fluorescein can be achieved using 3'-Fluorescein CPGs. Standard cleavage and deprotection with ammonium hydroxide liberates the fluorescein-labeled oligo when using any of these supports.
Calculators
Common stock solution preparation
Table 1. Volume of MeCN needed to reconstitute specific mass of 6-Fluorescein phosphoramidite to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 85.009 µL | 425.044 µL | 850.087 µL | 4.25 mL | 8.501 mL |
5 mM | 17.002 µL | 85.009 µL | 170.017 µL | 850.087 µL | 1.7 mL |
10 mM | 8.501 µL | 42.504 µL | 85.009 µL | 425.044 µL | 850.087 µL |
Molarity calculator
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Spectrum
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Product family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Correction Factor (280 nm) |
6-FAM phosphoramidite [5'-Fluorescein phosphoramidite] *CAS 204697-37-0* | 493 | 517 | 83000 | 0.178 |
6-TET phosphoramidite [5'-Tetrachlorofluorescein phosphoramidite] *CAS#: 877049-90-6* | 521 | 542 | 73000 | 0.13 |
6-HEX phosphoramidite [5'-Hexachlorofluorescein phosphoramidite] | 533 | 559 | 73000 | 0.15 |
6-JOE Phosphoramidite | 520 | 545 | 750001 | - |
Fluorescein-dT Phosphoramidite | 498 | 517 | 800001 | 0.35 |
6-VIC phosphoramidite | 526 | 543 | - | - |
References
View all 13 references: Citation Explorer
Sequence-specific, self-reporting hairpin inversion probes
Authors: Browne KA., undefined
Journal: J Am Chem Soc (2005): 1989
Authors: Browne KA., undefined
Journal: J Am Chem Soc (2005): 1989
Modified oligonucleotides containing lithocholic acid in their backbones: their enhanced cellular uptake and their mimicking of hairpin structures
Authors: Kim SJ, Bang EK, Kwon HJ, Shim JS, Kim BH.
Journal: Chembiochem (2004): 1517
Authors: Kim SJ, Bang EK, Kwon HJ, Shim JS, Kim BH.
Journal: Chembiochem (2004): 1517
Synthesis of HyBeacons and dual-labelled probes containing 2'-fluorescent groups for use in genetic analysis
Authors: Dobson N, McDowell DG, French DJ, Brown LJ, Mellor JM, Brown T.
Journal: Chem Commun (Camb) (2003): 1234
Authors: Dobson N, McDowell DG, French DJ, Brown LJ, Mellor JM, Brown T.
Journal: Chem Commun (Camb) (2003): 1234
Reduced aggregation and improved specificity of G-rich oligodeoxyribonucleotides containing pyrazolo[3,4-d]pyrimidine guanine bases
Authors: Kutyavin IV, Lokhov SG, Afonina IA, Dempcy R, Gall AA, Gorn VV, Lukhtanov E, Metcalf M, Mills A, Reed MW, S and ers S, Shishkina I, Vermeulen NM.
Journal: Nucleic Acids Res (2002): 4952
Authors: Kutyavin IV, Lokhov SG, Afonina IA, Dempcy R, Gall AA, Gorn VV, Lukhtanov E, Metcalf M, Mills A, Reed MW, S and ers S, Shishkina I, Vermeulen NM.
Journal: Nucleic Acids Res (2002): 4952
Diels--Alder bioconjugation of diene-modified oligonucleotides
Authors: Hill KW, Taunton-Rigby J, Carter JD, Kropp E, Vagle K, Pieken W, McGee DP, Husar GM, Leuck M, Anziano DJ, Sebesta DP.
Journal: J Org Chem (2001): 5352
Authors: Hill KW, Taunton-Rigby J, Carter JD, Kropp E, Vagle K, Pieken W, McGee DP, Husar GM, Leuck M, Anziano DJ, Sebesta DP.
Journal: J Org Chem (2001): 5352
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