North2South™ Biotin Random Prime DNA Labeling Kit

Catalog number: 17075

Thermo Scientific™  Related applications: Nucleic Acid Gel Electrophoresis & Blotting

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Description

Thermo Scientific North2South Random Prime DNA Biotinylation Kit uses random heptanucleotides, Klenow fragment and biotin-nucleotides to produce biotinylated DNA templates for DNA hybridization and detection methods.

Features of the North2South Random Prime DNA Biotinylation Kit:

• Works with as little as 100ng starting DNA template
• Exonuclease-free Klenow fragment in the DNA labeling kit produces higher yields
• Each reaction yields probe sufficient for approximately three (10 x 10 cm) blots

This random prime DNA labeling kit is based on the procedure of Feinberg and Vogelstein (Ref.4,5). Random heptanucleotides containing all possible sequences are annealed to a denatured DNA template. These act as primers for complementary strand synthesis by DNA Polymerase (Klenow fragment, 3'-5' exo-). Biotinylated dNTPs in the reaction mix are incorporated into the newly synthesized DNA. This protocol yields biotin-labeled DNA of high activity for use as probes in hybridization experiments such as Southern and Northern hybridizations. The resulting hybrids can be detected with streptavidin-horseradish peroxidase (HRP) and a chemiluminescent substrate kit such as North2South Chemiluminescent Hybridization and Detection Kit.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Product Size: 10 reactions

Contents & storage

Sufficient For: 10 biotin-labeling reactions (each with 100 ng DNA)
• Heptanucleotide Mix (5X), 100 µL
• Deoxynucleotide Mix (5X), 80 µL
• Reaction Buffer (10X), 50 µL
• DNA Polymerase Klenow Fragment (5U/µL), 10 µL
• Control DNA (250 ng/µL), 5 µL
• Ammonium Acetate (5M), 1 mL
• EDTA (0.5M, pH 8.0), 1 mL
• Nuclease-free Water, 1 mL
• Biotin-11-dUTP, 20 µL

Store all components at -20°C.

Documents

Manuals & protocols