In the T-RFLP method, PCR of the template DNA is performed using a primer having the 5' end labeled with a fluorescent molecule, and the resulting PCR product is digested with a restriction enzyme.
As only the terminal DNA fragment (T-RF) is detected by the fluorescent labeling, the features of restriction fragments, including fluorescence intensity, location, and number of peaks on the sample spectrum, can be analyzed by the profiling of microbial communities.