PREPARATION AND PURIFICATION OF DNA FROM INSECTS FOR AFLP ANALYSIS

Analysis of amplified fragment length polymorphism (AFLP) has the pote ntial to become a powerful new DNA fingerprinting technique for studyi ng genetic relationships and genetic diversity in arthropods. Since DN A of high quality is a crucial prerequisite for AFLP analysis we evalu ated the applicability of six protocols (one fast and four complex met hods with phenol-chloroform treatments as well as one CTAB-based metho d) for extracting DNA from insect material and three additional DNA pu rification steps. The most rapid DNA isolation method did not produce DNA suitable for AFLP analysis. Among four complex methods tested, two protocols resulted in comparatively low yields of DNA that was theref ore not used as template for AFLP analysis. The other two complex meth ods with phenol treatments and a CTAB-based DNA extraction protocol pr ovided DNA suitable for AFLP assay. An additional purification of the DNA using spermine precipitation revealed a few extra bands in an AFLP gel that were masked in unpurified DNA. Therefore spermine precipitat ion is recommended for AFLP templates.