RAPD Protocol
(Adapted from Williams et al., 1990*)
NOTES: This protocol has been optimized for a specific set of conditions and the following precautions should be noted.
DNA isolations must include a phenol extraction (Option A, p.2).
Always resuspend DNAs in sterile ddH2O (Sigmaís Cell Culture Water, Cat. # W-3500, provides an excellent standard).
Accurate quantification of DNA amounts is important; fluorometric measurements are usually more accurate than spectrophotometric ones at low concentrations.
The concentration of all DNAs should be adjusted to 5 ng/μl, and they should be aliquoted in appropriate amounts and frozen until needed.
Take note of the lot number of the available Taq enzyme and perform the necessary experiments to determine the optimum amounts of enzyme and of MgCl2 for best performance. Repeat these tests whenever you change to a new lot of enzyme.
Use filtered pipette tips for the preparation of all stocks of reagents included in these protocols, as well as for pipetting DNAs and primers.
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