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North2South Random Prime DNA Biotinylation Kit


A safe alternative to radiolabeling of oligonucleotide probes.

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.

Related Products:

North2South Chemiluminescent Hybridization and Detection Kit
DNA 3' End Biotinylation Kit
RNA 3' End Biotinylation Kit


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
17075 North2South Biotin Random Prime DNA Labeling Kit
Formulation: Multi-component kit with reagents for DNA labeling
Sufficient For: 10 biotin-labeling reactions (each with 100ng DNA)

Kit Contents:
Heptanucleotide Mix (5X), 100μL
Deoxynucleotide Mix (5X), 80μL
Reaction Buffer (10X), 50μL
DNA Polymerase Klenow Fragment (5U/μL), 10μL
Control DNA (250ng/μL), 5μL
Ammonium Acetate (5M), 1mL
EDTA (0.5M, pH 8.0), 1mL
Nuclease-free Water, 1mL
Biotin-11-UTP, 20μL
10-rxn kit Product Instructions for product #17075 North2South  Biotin Random Prime DNA Labeling Kit MSDS for product #17075 North2South  Biotin Random Prime DNA Labeling Kit Certificate of Analysis for product #17075 North2South  Biotin Random Prime DNA Labeling Kit $333.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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