MilliporeSigma
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11745816910

Roche

DIG-Nick Translation Mix

greener alternative

sufficient for 40 labeling reactions, pkg of 160 μL, suitable for hybridization

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Synonym(s):
nick translation

form

solution

Quality Level

usage

sufficient for 40 labeling reactions

packaging

pkg of 160 μL

manufacturer/tradename

Roche

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

technique(s)

hybridization: suitable

greener alternative category

storage temp.

−20°C

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This Item
117458089101174582491011277065910
DIG-Nick Translation Mix sufficient for 40 labeling reactions, pkg of 160 μL, suitable for hybridization

Roche

11745816910

DIG-Nick Translation Mix

Nick Translation Mix sufficient for 50 labeling reactions, pkg of 200 μL, solution

Roche

11745808910

Nick Translation Mix

DIG DNA Labeling Mix

Roche

11277065910

DIG DNA Labeling Mix

form

solution

form

solution

form

solution

form

solution

usage

sufficient for 40 labeling reactions

usage

sufficient for 50 labeling reactions

usage

sufficient for 40 labeling reactions

usage

sufficient for 25 standard reactions

packaging

pkg of 160 μL

packaging

pkg of 200 μL

packaging

pkg of 160 μL

packaging

pkg of 50 μL

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

greener alternative product characteristics

-

greener alternative product characteristics

-

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

General description

DIG-Nick Translation Mix is a convenient enzyme and nucleotide mixture for nick translation. It 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. Large plasmids, cosmids and polymerase chain reaction (PCR) fragments are all regularly used with this method and the DNA can be linearized or supercoiled. Individual templates produce consistent results in the standard 90-minutes reaction and result in an average probe length of 200 bp up to 500 bp. The molar ratio of digoxigenin (DIG)-11-deoxyuridine triphosphate (dUTP) to deoxythymidine triphosphate (dTTP) is adjusted to ensure that every 20th to 25th nucleotide in the newly synthesized DNA is modified with DIG. This density of haptens in the DNA yields the highest sensitivity in the immunological detection reaction.
We are committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product is designed as a safer chemical.  The DIG System was established as a sensitive and cost-effective alternative to radioactivity for the labeling and detection of nucleic acids. There are many available publications that prove the versatility of the DIG System, so use of radio-labeling is no longer the only option for labeling of DNA for hybridization.

Specificity

Heat inactivation: Stop the reaction by adding 1μl 0.5M EDTA (pH 8.0) and heating to 65°C for 10 minutes.

Application

DIG-Nick Translation Mix has been used for the generation of highly sensitive probes for in situ hybridization labeled with Digoxigenin-11-dUTP.
Note: The mix can also be used for filter hybridization techniques, however, for highly sensitive filter hybridization probes, we recommend that you use DIG-High Prime from Roche Applied Science.
For nonradioactive labeling of in situ probes with other haptens and fluorophores, Roche Applied Science offers the Biotin-Nick Translation Mix and the Nick Translation Mix (without nucleotides).

Quality

Function tested in a dot spot assay.

Principle

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.

Preparation Note

1 vial with 5x concentrated stabilized reaction buffer in 50% glycerol (v/v) and DNA Polymerase I, DNase I, 0.25mM dATP, 0.25mM dCTP, 0.25mM dGTP, 0.17mM dTTP and 0.08mM DIG-11-dUTP (alkali-stable).
Assay Time: 100 minutes
Sample Materials: Supercoiled and linearized plasmid DNA, Supercoiled and linearized cosmid DNA, Purified PCR products.
Note: Denaturing of the template before nick translation is not required.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

No data available

Flash Point(C)

No data available


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Piperidis N, et al.
International Sugar Journal, 1-9 (2013)
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Digoxigenin (DIG) Labeling Methods

Digoxigenin (DIG) labeling methods and kits for DNA and RNA DIG probes, random primed DNA labeling, nick translation labeling, 5’ and 3’ oligonucleotide end-labeling.

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