Application

Biotin-High Prime is used for the highly efficient random-primed labeling of DNA probes with Biotin-16-dUTP. Biotin-labeled probes are generated at high yield within one hour or after overnight incubation. Biotin-High Prime labeled probes are used in a variety of hybridization techniques: Southern blotsNorthern blotsDot/slot blotsScreening of gene librariesIn situ hybridizations

Features and Benefits

Contents5x concentrated random primer mix: 1U/µl Klenow polymerase, labeling grade, 1mM dATP, 1mM dCTP, 1mM dGTP, 0.65mM dTTP, 0.35mM Biotin-16-dUTP, in 50% (v/v) glycerol.

General description

Convenient enzyme and nucleotide mixture for rapid random-primed labeling of DNA with Biotin-16-dUTP. In this method, the complementary DNA strand of denatured DNA is synthesized by Klenow polymerase using the 3′-OH termini of the random oligonucleotides as primers.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Preparation Note

Assay Time: 80 minutesSample Materials DNA fragments of at least 100bp, Linearized plasmids, cosmid or λDNA, Supercoiled DNA,Or minimal amounts of DNA (10ng), e.g., DNA restriction fragments isolated from gels or in molten agarose can be used. Labeling efficiency: A standard labeling reaction with 1µg template yields 0.8µg biotin-labeled DNA after 1-hour, and 2µg after a 20-hour incubation at +37°C.

Specificity

Heat inactivation: Stop the reaction by adding 2 µl 0.2 M EDTA (pH 8.0) and/or by heating to 65 °C for 10 minutes.