North2South™ Biotin Random Prime DNA Labeling Kit uses random heptanucleotides, Klenow fragment and biotin-nucleotides to produce biotinylated DNA templates for DNA hybridization 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 x 100 m) blots
The probes are created when random heptanucleotides containing all possible sequences are annealed to a denatured DNA template
Once the labeled probes are ready for the membrane, use the chemiluminescent hybridization and detection kit (PI17097) to detect the streptavidin-HRP labeled probe. The kit combines an enhanced luminol substrate for horseradish peroxidase (HRP) with pre-mixed hybridization and blocking buffers to ensure consistent results with sensitivity equal to or greater than radioactive phosphorus techniques. The protocol is amenable to many formats and results are easily recorded by a 1 to 5 minute exposure to X-ray film or with a CCD camera.
These then act as primers for complementary strand synthesis by DNA polymerase. The product formulation uses biotinylated dCTP instead of dUTP to maximize the streptavidin-HRP binding and increase the sensitivity. Reactions can be done with as little as 100 ng of starting DNA template. The Klenow fragment in the labeling kit is exonuclease-free to produce higher yields. Each reaction will yield enough probe sample for three 10 x 10cm (315/16 x 315/16") blots.
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 ?L.
