Analysis of Loss of Heterozygosity in Circulating DNA

Analysis of genetic altera tion in circulating DNA can have clinical utility in predicting disease outcome. Loss of heterozygosity (LOH) of DNA microsatellites has been shown to occur commonly among all chromosomes in various cancers, such as melanoma, breast cancer, and lung cancer. In this protocol, we focused on the utility of LOH of microsatellite biomarkers for detection of analyzing circulating DNA. The protocol describes how PCR is performed on each patient’s paired DNA samples (normal lymphocyte DNA and serum DNA) using specific microsatellite biomarkers followed by post-PCR product analysis using capillary array electrophoresis (CAE). The utility of CAE is due to its digitalization and accuracy of the post-PCR product results.