6-carboxyfluorescein
Structural formula | |||||||||||||||||||
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General | |||||||||||||||||||
Surname | 6-carboxyfluorescein | ||||||||||||||||||
other names |
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Molecular formula | C 21 H 12 O 7 | ||||||||||||||||||
Brief description |
red solid |
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properties | |||||||||||||||||||
Molar mass | 376.32 g mol −1 | ||||||||||||||||||
Physical state |
firmly |
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Melting point |
> 300 ° C |
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solubility | |||||||||||||||||||
safety instructions | |||||||||||||||||||
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As far as possible and customary, SI units are used. Unless otherwise noted, the data given apply to standard conditions . |
6-Carboxyfluorescein ( 6-FAM ) is a fluorescent dye with an absorption wavelength of 495 nm and an emission wavelength of 517 nm . It can be classified both in the group of triphenylmethane dyes and xanthene dyes . 6-FAM is used in the sequencing of nucleic acids and in the marking of nucleotides .
The term FAM, which is frequently encountered, is incorrect, but takes into account the fact that the commercially available product consists of the two isomers 5-FAM and 6-FAM.
use
The phosphoramidite derivative 6-FAM-phosphoramidite is used for the quantitative analysis of nucleic acids using real-time PCR . In this case, in addition to the introduction of the phosphoramidite residue, the hydroxyl groups of 6-FAM at positions 3 'and 6' are esterified with dimethylpropanoic acid . What is the difference between the absorption and emission wavelengths of 6-FAM-phosphoramidite and 6-carboxyfluorescein.
Another known derivative of 6-carboxyfluorescein is its succinimidyl ester, which has the same application as phosphoramidite, but in contrast to this, like 6-FAM, has an absorption wavelength of 495 nm and an emission wavelength of 517 nm.
properties
In addition to the open carboxylic acid form, as with fluorescein , there is also the spiro - lactone form as a tautomer :
Individual evidence
- ↑ a b c Entry on 6-FAM at Anaspec, accessed on October 23, 2016.
- ↑ a b c Data sheet 6-Carboxyfluorescein, ~ 97% (HPLC) from Sigma-Aldrich , accessed on October 27, 2016 ( PDF ).
- ↑ entry to 6-carboxyfluorescein in the ChemIDplus database of the United States National Library of Medicine (NLM) .
- ↑ M. Adamczyk, JR Fishpaugh, KJ Heuser: Preparation of succinimidyl and pentafluorophenyl active esters of 5- and 6-carboxyfluorescein. In: Bioconjug. Chem. 1997, 8 (2), pp. 253-255. PMID 9095369 .
- ↑ Entry on 6-FAM, SE at Anaspec, accessed on October 23, 2016.
literature
- JW Brandis: Dye structure affects Taq DNA polymerase terminator selectivity. In: Nucleic Acids Res. 27 (8), 1999, pp. 1912-1918. PMID 10101201 . (PDF)
- JA Jordan et al: TaqMan-based detection of Trichomonas vaginalis DNA from female genital specimens. In: J. Clin. Microbiol. 39 (11), 2001, pp. 3819-3822. PMID 11682492 . PDF