Meeting Abstract
Amplified fragment length polymorphism (AFLP), a standard method for genetic fingerprinting, typically involves the use of radioactive isotopes. This technique can present financial and logistical challenges, as radioactive isotopes can be expensive and dangerous, especially in an undergraduate teaching laboratory. The goal of our research was to optimize a protocol for genetic fingerprinting different clone lines of pale anemones (Aiptasia pallida) that did not require the use of these radioactive isotopes. DNA from individual anemones was extracted and then digested with EcoRI and MseI restriction endonucleases. Adapters were then ligated to the cut sites, and DNA was amplified via nested PCR using EcoRI- and MseI-specific primers. In the second amplification step, EcoRI primers were labeled with fluorescent tags instead of radioactive isotopes. To analyze and compare the genetic fingerprints, the DNA was electrophoresed through 8% TBE polyacrylamide gels, which were photographed using a variable mode Typhoon Trio imager. The genetic fingerprints obtained using this method allowed us to distinguish genetically distinct clone lines from one another.
