NSH VIR Teleconference: Selection, Production and Characterization of Probes for In Situ Hybridization

Presented by Stephanie Villarreal, Merck

In situ hybridization has been established as a uniquely powerful tool for the study of gene expression in tissues, allowing both the visualization and quantification of gene expression at the regional and cellular level. While a highly sensitive and versatile technique, its value is dependent on the careful selection and characterization of probes. This presentation will focus on three critical aspects of probe selection. First, we will outline a strategy that can be used to select a probe that is unique to the gene sequence of interest. This section will also discuss ways to deal with similar unwanted genes such as other members of a super family of genes. Next we will discuss the benefits and drawbacks of ribonucleotide versus oligonucleotide probes and how their length can be used to enhance specificity. Finally, I will provide and overview of different methods used to label probes and provide examples generated using these different methods.

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