Non-isotopic procedure with silver staining of polyacrylamide gels in detection of genomic abnormalities in tumors using microsatellites

Microsatellite-based PCR (polymerase chain reaction) with silver staining of polyacrylamide gels as an alternative approach to detect genomic abnormalities in tumors has been developed. In experiments with a series of primers for microsatellite sequences located on human chromosome 5 we found that a 2- to 4-fold increase or decrease in the ratio of amount of allelic microsatellite sequence can be reliably assessed from the relative intensity of corresponding PCR products. Results of the studies of renal carcinoma cell lines carrying increased or decreased numbers of specific allelic markers on chromosome 5 assessed by cytogenetics, Southern blotting-restriction fragment length polymorphism (using 32P as a labelled compounds) and microsatellite analyses were consistent. Microsatellite studies performed on samples obtained from formalin-fixed paraffin-embedded archival material allowed detection of genomic abnormalities of chromosome 5 in ten primary renal cell carcinomas. (author). 35 refs, 4 figs, 4 tabs