Chapter 35
PRINS: Hybridization and Primed in Situ labeling
in One Step
JORN KOCH
Introduction
PRINS (PRimed IN Situ labeling) is a technique which is primarily used for
the detection of tandernly repeated target sequences in chromosomes and
nuclei ("in situ") by oligonucleotide probes. Such sequences can be detected down to a target size of a few hundred bases with a simple short
procedure rendering virtually 100% of potential targets labeled.
The PRINS concept was introduced by the author of this text (Koch et
al. 1989) for the study of minute details in centromere DNA in situ; details
so small that they would be difficult to study by in situ hybridization (in
casu sequence variation among 16-bp target sequences within an array of
repeated sequences). The extent to which the technique can discriminate
among closely related sequences has more recently also been illustrated by
Pellestor and coworkers (1994,1995), and by us (Koch et al. 1995), both
discriminating between the alpha satellite DNA of chromosome 13,
and that of chromosome 21, on the basis of a single base difference. A
further advantage of the PRINS technique is that the probes used are
very small, and thus penetrate easily to the target, even if this is in a
very tight formation. This feature of the technique is being exploited
by the group of Franck Pellestor to enable efficient staining of target sequences in sperm cells (e.g. see Pellestor et al. 1997).
More recently, a new version of the PRINS technique, the "dideoxyPRINS", has been developed to enable highly sensitive, quantitative staining of simple repeat DNA, including telomeric repeats and trinucleotide
repeats. The dideoxy-PRINS can be understood from the basic PRINS
principle outlined below (Fig. 1). In PRINS, the probe is unlabeled,
and thus does not itself label the target binding it. Labeling is only obJorn Koch, Cancercytogenetics Laboratory, Department of Hematology, rage Hansens
Gade 2, Aarhus C, 8000, Denmark (phone +45-8949-7398; fax +45-8949-7399;
e-mail Jorn.Koch@aas.auh.dkorjornkoch@hotmail.com)
PROTOCOL
PRINS: Hybridization and Primed in Situ labeling
in One Step
JORN KOCH
Introduction
PRINS (PRimed IN Situ labeling) is a technique which is primarily used for
the detection of tandernly repeated target sequences in chromosomes and
nuclei ("in situ") by oligonucleotide probes. Such sequences can be detected down to a target size of a few hundred bases with a simple short
procedure rendering virtually 100% of potential targets labeled.
The PRINS concept was introduced by the author of this text (Koch et
al. 1989) for the study of minute details in centromere DNA in situ; details
so small that they would be difficult to study by in situ hybridization (in
casu sequence variation among 16-bp target sequences within an array of
repeated sequences). The extent to which the technique can discriminate
among closely related sequences has more recently also been illustrated by
Pellestor and coworkers (1994,1995), and by us (Koch et al. 1995), both
discriminating between the alpha satellite DNA of chromosome 13,
and that of chromosome 21, on the basis of a single base difference. A
further advantage of the PRINS technique is that the probes used are
very small, and thus penetrate easily to the target, even if this is in a
very tight formation. This feature of the technique is being exploited
by the group of Franck Pellestor to enable efficient staining of target sequences in sperm cells (e.g. see Pellestor et al. 1997).
More recently, a new version of the PRINS technique, the "dideoxyPRINS", has been developed to enable highly sensitive, quantitative staining of simple repeat DNA, including telomeric repeats and trinucleotide
repeats. The dideoxy-PRINS can be understood from the basic PRINS
principle outlined below (Fig. 1). In PRINS, the probe is unlabeled,
and thus does not itself label the target binding it. Labeling is only obJorn Koch, Cancercytogenetics Laboratory, Department of Hematology, rage Hansens
Gade 2, Aarhus C, 8000, Denmark (phone +45-8949-7398; fax +45-8949-7399;
e-mail Jorn.Koch@aas.auh.dkorjornkoch@hotmail.com)
PROTOCOL