6-carboxyfluorescein

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Structural formula
Structural formula of 6-carboxyfluorescine
General
Surname 6-carboxyfluorescein
other names
  • 3 ′, 6′-dihydroxy-1-oxospiro [2-benzofuran-3,9′-xanthene] -5-carboxylic acid ( IUPAC )
  • 3 ', 6'-Dihydroxy-3-oxospiro [isobenzofuran-1 (3 H ), 9' - (9 H ) xanthene] -6-carboxylic acid
  • 6-FAM
  • FLUOS
Molecular formula C 21 H 12 O 7
Brief description

red solid

External identifiers / databases
CAS number 3301-79-9
EC number 636-503-2
ECHA InfoCard 100.164.295
PubChem 76806
ChemSpider 69262
Wikidata Q245486
properties
Molar mass 376.32 g mol −1
Physical state

firmly

Melting point

> 300 ° C

solubility

soluble in DMF or DMSO

safety instructions
GHS labeling of hazardous substances
no GHS pictograms
H and P phrases H: no H-phrases
P: no P-phrases
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 :

Tautomerism

Individual evidence

  1. ↑ a b c Entry on 6-FAM at Anaspec, accessed on October 23, 2016.
  2. ↑ a b c Data sheet 6-Carboxyfluorescein, ~ 97% (HPLC) from Sigma-Aldrich , accessed on October 27, 2016 ( PDF ).
  3. ↑ entry to 6-carboxyfluorescein in the ChemIDplus database of the United States National Library of Medicine (NLM) .
  4. ↑ 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 .
  5. ↑ 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