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Fluorescein-12-dUTP *1 mM in Tris Buffer (pH 7.5)* *CAS 214154-36-6*

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Physical properties
Molecular weight1060.62
SolventWater
Spectral properties
Absorbance (nm)487
Correction Factor (260 nm)0.32
Correction Factor (280 nm)0.35
Extinction coefficient (cm -1 M -1)800001
Excitation (nm)498
Emission (nm)517
Quantum yield0.79001, 0.952
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC41116134

OverviewpdfSDSpdfProtocol


CAS
214154-36-6
Molecular weight
1060.62
Absorbance (nm)
487
Correction Factor (260 nm)
0.32
Correction Factor (280 nm)
0.35
Extinction coefficient (cm -1 M -1)
800001
Excitation (nm)
498
Emission (nm)
517
Quantum yield
0.79001, 0.952
The dye-modified deoxyuridine 5'-triphosphates (such as aminoallyl-dUTP) can be used to produce dye-containing DNA by conventional enzymatic incorporation methods such as reverse transcription, nick translation, random primed labeling, or PCR. This enzymatic fluorescence labeling method is widely used for both FISH probes and microarray-based experiments. This fluorescein-dUTP conjugate can be used as a green fluorescence color with Spectrum Green™ filter set (Spectrum Green™ is the trademark of Vysis).

Example protocol


AT A GLANCE

Important notes

Best stored at -80 °C, it can be diluted 10 folds in TE Buffer (10mM Tris-HCl + 1mM EDTA (pH=7.5)) for convenient pipetting. Expiration date is six months from the date of receipt.

SAMPLE EXPERIMENTAL PROTOCOL

Table:1. The following instruction is recommended as a starting point for labeling ~1µg dsDNA, optimum labeling conditions may vary for different cases. 

Reagents

Final Concentration for 20µL reaction volume

DNA

0.05 µg/µL

DNA Polymerase

25 ~ 50U/mL

Fluorescein-dUTP

10 ~ 100 µM

Reaction Buffer

10 mM Tris-HCl (pH = 7.5) contains the following components:
1 mM EDTA
5 mM NaCl
0.1 mM DTT
1 mM dATP + 1 mM dCTP+ 1 mM dGTP+ 1 mM dTTP

ddH2O

Adjust volume as need to make 20 µL reaction volume

Calculators


Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of Fluorescein-12-dUTP *1 mM in Tris Buffer (pH 7.5)* *CAS 214154-36-6* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM94.284 µL471.422 µL942.845 µL4.714 mL9.428 mL
5 mM18.857 µL94.284 µL188.569 µL942.845 µL1.886 mL
10 mM9.428 µL47.142 µL94.284 µL471.422 µL942.845 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
/=x=

Spectrum


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spectrum

Spectral properties

Absorbance (nm)487
Correction Factor (260 nm)0.32
Correction Factor (280 nm)0.35
Extinction coefficient (cm -1 M -1)800001
Excitation (nm)498
Emission (nm)517
Quantum yield0.79001, 0.952

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
Fluorescein-12-dCTP *1 mM in Tris Buffer (pH 7.5)* 4985178000010.79001, 0.9520.320.35
Fluorescein-12-dATP *1 mM in Tris Buffer (pH 7.5)* 4985178000010.79001, 0.9520.320.35
Fluorescein-12-dGTP *1 mM in Tris Buffer (pH 7.5)* 4985178000010.79001, 0.9520.320.35

Images


References


View all 9 references: Citation Explorer
Fluorescent DNA hybridization probe preparation using amine modification and reactive dye coupling
Authors: Cox WG, Singer VL.
Journal: Biotechniques (2004): 114
A visual DNA chip for simultaneous detection of hepatitis B virus, hepatitis C virus and human immunodeficiency virus type-1
Authors: Wen JK, Zhang XE, Cheng Z, Liu H, Zhou YF, Zhang ZP, Yang JH, Deng JY.
Journal: Biosens Bioelectron (2004): 685
Aminomodified nucleobases: functionalized nucleoside triphosphates applicable for SELEX
Authors: Schoetzau T, Langner J, Moyroud E, Roehl I, Vonhoff S, Klussmann S.
Journal: Bioconjug Chem (2003): 919
Simple method for preparation of fluor/hapten-labeled dUTP
Authors: Nimmakayalu M, Henegariu O, Ward DC, Bray-Ward P.
Journal: Biotechniques (2000): 518
Topology of yeast RNA polymerase II subunits in transcription elongation complexes studied by photoaffinity cross-linking
Authors: Wooddell CI, Burgess RR.
Journal: Biochemistry (2000): 13405
Quantitative analysis of polymerase chain reaction products using biotinylated dUTP incorporation
Authors: Duplaa C, Couffinhal T, Labat L, Moreau C, Lamaziere JM, Bonnet J.
Journal: Anal Biochem (1993): 229
A non-radioisotopic reverse transcriptase assay using biotin-11-deoxyuridinetriphosphate on primer-immobilized microtiter plates
Authors: Urabe T, Sano K, Tanno M, Mizoguchi J, Otani M, Lee MH, Takasaki T, Kusakabe H, Imagawa DT, Nakai M.
Journal: J Virol Methods (1992): 145
Affinity isolation of active murine erythroleukemia cell chromatin: uniform distribution of ubiquitinated histone H2A between active and inactive fractions
Authors: Dawson BA, Herman T, Haas AL, Lough J.
Journal: J Cell Biochem (1991): 166
Non-radioactive labeling and detection of nucleic acids. IV. Synthesis and properties of digoxigenin-modified 2'-deoxyuridine-5'-triphosphates and a photoactivatable analog of digoxigenin (photodigoxigenin)
Authors: Muhlegger K, Huber E, von der Eltz H, Ruger R, Kessler C.
Journal: Biol Chem Hoppe Seyler (1990): 953