CHEMILUMINESCENT DETECTION OF BLOTTED PCR PRODUCTS (CB-PCR) OF 2 CAG DYNAMIC MUTATIONS (HUNTINGTONS-DISEASE AND SPINOCEREBELLAR ATAXIA TYPE-1)

We have used a non-isotopic PCR assay based on the chemiluminescent de tection of blotted PCR products (CB-PCR) for two dynamic mutation dise ases (Huntington's disease and spinocerebellar ataxia type 1). This gi ves an accurate sizing of alleles and permits a rapid analysis of at r isk persons. The system involves PCR of the samples, separation of all eles on polyacrylamide gels, Southern blotting, and hybridisation with specific primers 3' labelled with fluorescein (F1)-dUTP as probes. CB -PCR retains the isotopic sensitivity for accurate allele determinatio n, avoids isotopic manipulation, and provides the advantages of safety , long term storage of probes, and recycling of hybridisation solution s.