The introduction of primed in situ labeling (PRINS) into plant cytogenetics provided a novel means for fast and highly specific visualization of DNA sequences on chromosomes and in interphase nuclei. Although the technique does not reach the sensitivity of fluorescence in situ hybridization that is needed for detection of single-copy targets, it is superior in its speed and simplicity. Thus, the main applications of PRINS include fluorescent labeling of repeated DNA sequences, such as ribosomal DNA and satellite repeats, which are used to discriminate individual chromosome types within karyotypes and to assess the purity of chromosome fractions separated by flow-sorting. Recently, an application of PRINS for the discrimination of sequence subfamilies of satellite repeats has been developed that takes advantage of the sensitivity of Taq polymerase to mismatches between 3'-end of the primer and the template sequences. This approach allows the distinguishing of sequences that differ in only a few nucleotides and has proved to be valuable for studies of satellite DNA evolution in plants.
Key Words: Plant chromosomes; repetitive sequences; satellite DNA; flow cytometric sorting.
Was this article helpful?