Principles and Applications of PRINS in Cytogenetics

AbstractPRINS (PRimed IN Situ labeling) is a technique that traditionally has been used for the detection of tandemly repeated target sequences in chromosomes and nuclei (“in situ”) by oligonucleotide probes. With a simple short procedure rendering virtually 100% of potential targets labeled, such sequences can be detected down to a target size of a few hundred bases. More recently it has been found that single‐copy sequences can also be detected at high sensitivity, if signals are enhanced with the Tyramide system. This unit provides protocols for basic PRINS or dideoxy‐PRINS (ddPRINS), and for combinatorial labeling (combining PRINS with FISH). These protocols are also applicable after immunostaining, provided that the antibody has been “secured” at the site by fixation in (para)formaldehyde.