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 dige stion 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 i s accomplished in one reaction vs. two. PCR products are resolved in agaros e-Synergel instead of polyacrylamide and are visualized by ethidium bromide staining rather than by autoradiography of labeled primers. Using the modi fied procedure for honey bee DNA, high reproducibility of the band patterns of PCR products and low sensitivity to the amplification conditions were s een. Analysis of honey bee DNA revealed considerable genetic variability wi thin and between African and European bee samples. African- and European-sp ecific fragments were found.