11745808910
Roche
Nick Translation Mix
sufficient for 50 labeling reactions, pkg of 200 μL, solution
Synonym(e):
nick translation
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About This Item
UNSPSC-Code:
41105500
Empfohlene Produkte
Form
solution
Qualitätsniveau
Verwendung
sufficient for 50 labeling reactions
Verpackung
pkg of 200 μL
Hersteller/Markenname
Roche
Methode(n)
nucleic acid labeling: suitable
Farbe
colorless
pH-Wert
~7.5 (68 °F)
Löslichkeit
water: miscible
Eignung
suitable for fluorescent labeling techniques
suitable for molecular biology
Anwendung(en)
genomic analysis
life science and biopharma
Lagertemp.
−20°C
Allgemeine Beschreibung
Optimized enzyme mixture: Nick translation utilizes a combination of DNase and DNA Polymerase to nick one strand of the DNA helix, then incorporates labeled nucleotides as the polymerase examines, or "proofreads" the nicked site.
Individual templates produce consistent results in the standard 90-minutes reaction, and result in an average probe length of 200 base pairs up to 500 base pairs.
Assay Time: 100 minutes
Sample Materials
Individual templates produce consistent results in the standard 90-minutes reaction, and result in an average probe length of 200 base pairs up to 500 base pairs.
Assay Time: 100 minutes
Sample Materials
- Supercoiled and linearized plasmid DNA
- Supercoiled and linearized cosmid DNA
- Purified PCR products
Spezifität
Heat inactivation: Stop the reaction by adding 1 μl 0.5 M EDTA (pH 8.0) and heating to 65 °C for 10 minutes.
Anwendung
For generation of highly sensitive probes for fluorescence in situ hybridization (FISH).
The Nick Translation Mix is designed for direct fluorophore-labeling of in situ probes. Fluorescein-12-dUTP and Tetramethyl-Rhodamine-5-dUTP from Roche Applied Science or other commercially available fluorophor-labeled nucleotides can be combined with the Nick Translation Mix. Direct fluorophore-labeled in situ probes are used for the detection of multi copy or very large hybridization targets on metaphase chromsomes or interphase nuclei.
For a standard labeling reaction using 1 μg template in 20 μl total reaction volume, 4 μl of 5x concentrated fluorophore labeling mix are required.
The Nick Translation Mix is designed for direct fluorophore-labeling of in situ probes. Fluorescein-12-dUTP and Tetramethyl-Rhodamine-5-dUTP from Roche Applied Science or other commercially available fluorophor-labeled nucleotides can be combined with the Nick Translation Mix. Direct fluorophore-labeled in situ probes are used for the detection of multi copy or very large hybridization targets on metaphase chromsomes or interphase nuclei.
For a standard labeling reaction using 1 μg template in 20 μl total reaction volume, 4 μl of 5x concentrated fluorophore labeling mix are required.
Komponenten
Contents
1 vial with 5x concentrated solution, stabilized reaction buffer in 50% glycerol (v/v), DNA Polymerase I and DNase I.
1 vial with 5x concentrated solution, stabilized reaction buffer in 50% glycerol (v/v), DNA Polymerase I and DNase I.
Qualität
Function tested in dot spot assay.
Prinzip
The nick translation method is based on the ability of DNase I to introduce randomly distributed nicks into DNA at low enzyme concentrations in the presence of MgCl2.
E. coli DNA Polymerase I synthesizes DNA complementary to the intact strand in a 5′?3′ direction using the 3′-OH termini of the nick as a primer. The 5′?3′ exonucleolytic activity of DNA polymerase I simultaneously removes nucleotides in the direction of synthesis. The polymerase activity sequentially replaces the removed nucleotides with isotope-labeled or hapten-labeled deoxyribonucleoside triphosphates. At low temperature (+15°C), the unlabeled DNA in the reaction is thus replaced by newly synthesized labeled DNA.
E. coli DNA Polymerase I synthesizes DNA complementary to the intact strand in a 5′?3′ direction using the 3′-OH termini of the nick as a primer. The 5′?3′ exonucleolytic activity of DNA polymerase I simultaneously removes nucleotides in the direction of synthesis. The polymerase activity sequentially replaces the removed nucleotides with isotope-labeled or hapten-labeled deoxyribonucleoside triphosphates. At low temperature (+15°C), the unlabeled DNA in the reaction is thus replaced by newly synthesized labeled DNA.
Lagerung und Haltbarkeit
Avoid repeated freezing and thawing.
Sonstige Hinweise
For life science research only. Not for use in diagnostic procedures.
Denaturing of the template before nick translation is not required.
Denaturing of the template before nick translation is not required.
Lagerklassenschlüssel
12 - Non Combustible Liquids
WGK
WGK 1
Flammpunkt (°F)
does not flash
Flammpunkt (°C)
does not flash
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