Spectral Karyotyping to Study Chromosome Abnormalities in Humans and Mice with Polycystic Kidney Disease

The aim of this procedure is to label chromosomes with paint probes to identify chromosome aberrations and segregation defects in mice and humans with polycystic kidney disease. This is accomplished by first harvesting cells from humans on mice and treating with smid, followed by fixing and preparing metaphase spreads on glass slides. Next, the slide is pretreated to remove extra RNA and cytoplasm which might interfere with the hybridization.

This is done by digestion with RNAs and pepsin respectively. Then chromosome and probe denaturation is performed followed by hybridization to label chromosomes. The final step is detection of the probe with fluorescent dyes.