Modification of the AFLP protocol applied to honey bee (Apis mellifera L.) DNA.

The established amplified fragment-length polymorphism (AFLP) protocol was simplified and optimized for honey bee DNA (Apis mellifera L.). Compared to the original method, the following simplifications were made: (i) the digestion of DNA and ligation of the adapters are performed in one reaction vs. two, (ii) one restriction enzyme is used vs. two and (iii) amplification is accomplished in one reaction vs. two. PCR products are resolved in agarose-Synergel instead of polyacrylamide and are visualized by ethidium bromide staining rather than by autoradiography of labeled primers. Using the modified procedure for honey bee DNA, high reproducibility of the band patterns of PCR products and low sensitivity to the amplification conditions were seen. Analysis of honey bee DNA revealed considerable genetic variability within and between African and European bee samples. African- and European-specific fragments were found.