North2South™ Biotin Random Prime DNA Labeling Kit uses random heptanucleotides, Klenow fragment and biotin-nucleotides to produce biotinylated DNA templates for DNA hybridisation and detection methods.
- Works with as little as 100 ng starting DNA template
- Exonuclease-free Klenow fragment in the DNA labeling kit produces higher yields
- Each reaction yields probe sufficient for approximately three (100×100 m) blots
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 synthesised DNA. This protocol yields biotin-labeled DNA of high activity for use as probes in hybridisation experiments such as Southern and Northern hybridisations. The resulting hybrids can be detected with streptavidin-horseradish peroxidase (HRP) and a chemiluminescent substrate kit.
Delivery information: Kit includes 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.
