North2South Random Prime DNA Biotinylation Kit

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.

Highlights:

• Works with as little as 100ng starting DNA template
• Exonuclease-free Klenow fragment in the DNA labeling kit produces higher yields
• Each reaction yields probe sufficient for approximately three (10 x 10cm) blots

Product Details:

This random prime DNA 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. The resulting hybrids can be detected with streptavidin-horseradish peroxidase (HRP) and a chemiluminescent substrate kit such as North2South Chemiluminescent Hybridization and Detection Kit.

References:

1. Hirsch, J. and Henry, S. (1986). Expression of the Saccharomyces cerevisiae inositol-1-phosphate synthase (INO1) gene is regulated by factors that affect phospholipid synthesis. Mol. Cell. Biol. 6, 3320-3328.
2. Hoffman, C. and Winston, F. (1987). A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57, 267-272.
3. Sambrook, J., Fritsch, E. and Maniatis, T. (eds.). Molecular Cloning, 2nd Ed. (1989). Cold Spring Harbor Press. Plainview, New York. (Product # 17946)
4. Feinberg, A.P. and Vogelstein, B. (1983). Anal. Biochem. 132, 6.
5. Feinberg, A.P. and Vogelstein, B. (1983). Anal. Biochem. 137, 266.

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