PI17075
Thermo Scientific™ North2South™ Biotin Random Prime DNA Labeling Kit
Manufacturer: Thermo Scientific™
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Content And 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 μLStore all components at -20°C.
Labeling Target
DNA
Product Type
DNA Labeling Kit
Format
Kit
Product Line
North2South™
Quantity
10 Reactions
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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
- 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