DIG DNA LABELING MIXTURE

General description

DIG DNA Labeling Mix is an easy-to-use labeling mixture for rapid random-primed labeling with digoxigenin (DIG)-11-deoxyuridine triphosphate (dUTP). DIG-dUTP is incorporated every 20 to 25 nucleotides in the newly synthesized DNA. This density of haptens in the DNA yields the highest sensitivity in the 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.

Application

The DIG DNA labeling mix has been used in a variety of hybridization techniques including:

• Southern blots
• northern blots
• dot blots
• screening of gene libraries
• in situ hybridizations

Features and Benefits

Contents
10x concentrated dNTP labeling mixture: 1mM dATP, 1mM dCTP, 1mM dGTP, 0.65mM dTTP, 0.35mM DIG-dUTP, alkali-labile, pH 7.5 (+20°C)

Assay Time: Labeling: 1 hour to O/N
Labeling efficiency: With 1μg DNA per assay, approx. 10% of the nucleotides are incorporated into about 250ng of newly synthesized labeled DNA within 1 hour and approx. 30% of the nucleotides into about 750ng after 20 hours.

Quality

Function-tested in the DIG DNA Labeling Kit and in the DIG Nucleic Acid Detection Kit.

Principle

DIG-labeled DNA probes are generated according to the random-primed labeling method which is based on the hybridization of random oligonucleotides to the denatured DNA template. The complementary DNA strand is synthesized by Klenow enzyme which uses the 3′-OH termini of the random oligonucleotides as primers and a mixture of deoxyribonucleotides containing DIG-11-dUTP, alkali-labile, for elongation. DIG-dUTP is incorporated every 20 to 25 nucleotides in the newly synthesized DNA. This density of haptens in the DNA yields the highest sensitivity in the detection reaction.

Preparation Note

Sample Materials
As template for the labeling reaction

• DNA fragments of at least 100bp
• Linearized plasmids, cosmid or λDNA,
• Supercoiled DNA,
• Or minimal amounts of DNA (10ng), e.g., DNA restriction fragments isolated from low melting point agarose can be used.

Other Notes

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