The North2South Random Prime 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.
| Sufficient reagents for 10 reactions.** | |
| Kit contains: | |
| 5X Heptanucleotide Mix | 100 µl |
| 5X Deoxynucleotide Mix | 80 µl |
| 10X Reaction Buffer | 50 µl |
| Klenow DNA Polymerase | 10 µl |
| Non-biotinylated control DNA | 5 µl |
| 5M Ammonium Acetate | 1 ml |
| 0.5M EDTA | 1 ml |
| Nuclease-free Water | 1 ml |
| Biotin-11-dUTP | 20 µl |
