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Thermo Scientific™ North2South™ Biotin Random Prime DNA Labeling Kit

Produce biotinylated DNA templates for DNA hybridization and detection methods using random heptanucleotides, Klenow fragment and biotin-nucleotides.

Manufacturer:  Thermo Scientific™ 17075

Catalog No. PI17075


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Description

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.

Random heptanucleotides containing all possible sequences are annealed to a denatured DNA template. These act as primers for complementary strand synthesis by DNA polymerase. 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).

Highlights:

  • Ultimate sensitivity - labels with biotinylated dUTP, maximizing streptavidin-HRP binding and increasing sensitivity
  • Works with as little as 100ng starting DNA template
  • Exonuclease-free Klenow fragment produces higher yields
  • Each reaction yields probe sufficient for approximately three (10 x 10cm) blots
Specifications

Specifications

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.
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