Detection of loss of heterozygosity by a tube-based assay

Here we report a tube format for detecting loss of heterozygosity in human genomic DNA. This method is based on the detection of a polymorphic restriction site in the selected gene by tube support analysis. After a PCR, amplification products were hybridized to an 125I-labeled oligonucleotide probe and affinity-collected on a tube. The captured hybrids were then subjected to an enzymatic digestion, and the remaining bound radioactivity was measured. The usefulness of this technique is illustrated by analyzing the p53 gene LOH in human colorectal samples. This development, similar to immunoanalysis tests, eliminates the electrophoresis step and makes the results easy to study.