Storage: −20°C
UNSPSC Code: 41106000
RIDADR: NONH for all modes of transport
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.
Application:
• The DIG DNA Labeling Mix is used for random primed DNA labeling with Digoxigenin-11-dUTP, alkali-labile. DIG-labeled probes are used in a variety of hybridization techniques: Southern blots
• Northern blots
• Dot blots
• Screening of gene libraries
• In situ hybridizations
General description:
Easy-to-use labeling mixture for rapid random-primed labeling with Digoxigenin-11-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.
Other Notes:
For life science research only. Not for use in diagnostic procedures.
Quality:
Function-tested in the DIG DNA Labeling Kit and in the DIG Nucleic Acid Detection Kit.
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.
Note: Linear DNA is labeled more efficiently than circular and supercoiled DNA.
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.