Thermo Scientific™ North2South™ Biotin Random Prime Labeling Kit
Produce biotinylated DNA templates for DNA hybridization and detection methods using random heptanucleotides, Klenow fragment and biotin-nucleotides.
Manufacturer: Thermo Scientific™ PI17075
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).
- 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
|Non-radioactive Southern blotting|
|5X Heptanucleotide Mix, 100µL;5X Deoxynucleotide Mix, 80µL;10X Reaction Buffer, 50µL;Klenow DNA Polymerase, 10µL;Control DNA, 5µL;5 M Ammonium Acetate, 1mL;0.5 M EDTA, pH 8.0, 1mL;Nuclease-free Water, 1mL;Biotin-11-UTP, 20µL|
|North2South DNA Probe Biotinylation Kit|
Multi-component kit with reagents for DNA labeling
|10 biotin-labeling reactions (each with 100ng DNA)|