DNA-LAB/DET-KIT NONRADIOACTIVE

General description

DIG DNA Labeling and Detection Kit is a convenient kit for random-primed labeling of DNA with digoxigenin-deoxyuridine triphosphate (dUTP), alkali-labile and color detection of hybrids by enzyme immunoassay. In this method, the complementary strand of denatured DNA is synthesized by Klenow polymerase using the 3′-OH termini of random oligonucleotides as primers.

Contents:

• Unlabeled Control DNA 1, 100 μg/ml
• Unlabeled Control DNA 2, 200 μg/ml
• DNA Dilution Buffer
• DIG-labeled Control DNA, 5.2 μg/ml
• 10x Hexanucleotide Mix
• 10x dNTP Labeling Mixture
• Klenow Enzyme, Labeling grade, 2 U/μl
• Anti-digoxigenin-AP-conjugate, 750 U/ml
• NBT/BCIP Concentrated Stock Solution
• Blocking Reagent

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

Sensitivity and specificity: The standard labeling reaction with 1 μg template will give a yield of 260 ng DIG-labeled DNA after a 1-hour incubation at +37°C, or 780 ng after a 20-hour incubation. The sensitivity of DIG detection depends on the concentration of the DIG-labeled probe in the hybridization reaction, and the duration of the color reaction.

Application

DIG DNA Labeling and Detection Kit has been used in a variety of hybridization techniques:

• Southern blots
• Northern blots
• Dot blots
• Colony and plaque screening

DIG-labeled hybrids are detected with an anti-DIG-alkaline phosphatase conjugate and the substrates NBT (nitroblue tetrazolium salt) and BCIP (5-bromo-4-chloro-3-indolyl phosphate, toluidinium salt), which give a light blue precipitate.

Packaging

1 kit containing 10 components.

Preparation Note

Working concentration: Dilution of Antibody 1:5,000

Other Notes

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