A modified hemi-nested primer system was used to amplify the CAG repeat region of the Huntington's gene. Details will be supplied. A total of 98 single fibroblasts were analyzed. Amplification of alleles from non-affected human blastomeres will be presented. Preliminary results show that single cell analysis of a triple base pair repeat disease is possible without the use of fluorescence technology.
ADO remains a significant problem when using a single cell.
However, the chance of misdiagnosis can be reduced significantly by using two cells for analysis when available. Optimization of the PCR reaction remains critical to the success of triple base pair amplification and new technology may also help in the analysis of amplified products.