Fluorescein-12-dUTP *1 mM in TE Buffer (pH 7.5)* *CAS 214154-36-6*
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: |
ddH2O |
Adjust volume as need to make 20 µL reaction volume |
Spectrum
Open in Advanced Spectrum Viewer
Product family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
Fluorescein-12-dCTP *1 mM in Tris Buffer (pH 7.5)* | 498 | 517 | 800001 | 0.79001, 0.952 | 0.32 | 0.35 |
Fluorescein-12-dATP *1 mM in Tris Buffer (pH 7.5)* | 498 | 517 | 800001 | 0.79001, 0.952 | 0.32 | 0.35 |
Fluorescein-12-dGTP *1 mM in Tris Buffer (pH 7.5)* | 498 | 517 | 800001 | 0.79001, 0.952 | 0.32 | 0.35 |
References
View all 9 references: Citation Explorer
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
Authors: Wen JK, Zhang XE, Cheng Z, Liu H, Zhou YF, Zhang ZP, Yang JH, Deng JY.
Journal: Biosens Bioelectron (2004): 685
Fluorescent DNA hybridization probe preparation using amine modification and reactive dye coupling
Authors: Cox WG, Singer VL.
Journal: Biotechniques (2004): 114
Authors: Cox WG, Singer VL.
Journal: Biotechniques (2004): 114
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
Authors: Schoetzau T, Langner J, Moyroud E, Roehl I, Vonhoff S, Klussmann S.
Journal: Bioconjug Chem (2003): 919
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
Authors: Wooddell CI, Burgess RR.
Journal: Biochemistry (2000): 13405
Simple method for preparation of fluor/hapten-labeled dUTP
Authors: Nimmakayalu M, Henegariu O, Ward DC, Bray-Ward P.
Journal: Biotechniques (2000): 518
Authors: Nimmakayalu M, Henegariu O, Ward DC, Bray-Ward P.
Journal: Biotechniques (2000): 518
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