This document provides instructions for using the Prime-It II Random Primer Labeling Kit. The product is intended for in vitro use only and comes with a limited warranty.
The document describes the Stratagene (Agilent) Prime-It II Random Primer Labeling Kit, designed to produce high-specific-activity DNA probes rapidly. The method utilizes a 3' exonuclease-deficient mutant of Klenow fragment and random nonamer primers to incorporate radiolabeled nucleotides into probe fragments in as little as two minutes.
The manual provides information on materials provided, storage conditions, and additional reagents required such as [a-³²P]dCTP or [a-³²P]dATP. It includes a quick-reference protocol for the labeling reaction, details on the time course of incorporation, sensitivity data, probe fragment lengths, and specific activities achieved with various template sizes.
| Template Size Range | 400 bp to 10 kb |
|---|---|
| Reaction Time for High Activity | 2 minutes |
| Nucleotide Incorporation Rate (2 min) | 59% |
| Minimum Detectable Target DNA | 1.0 pg |
The kit makes it possible to produce high-specific-activity DNA probes within two minutes.
The reaction should be incubated at 37-40°C.