The Future

It is always difficult to forecast the future, but several predictions can be made for the next few years. First, real-time PCR will become widely established in the analysis of methy-lation. Real-time PCR allows faster analysis and higher throughput because there are no further manipulations after the PCR step. However, the principal advantage is that molecular detection of low levels of disease benefits from the absence of post sample processing that real-time PCR analysis allows. Although stringent precautions are taken to eliminate cross-contamination, not having to remove the PCR products from the tube for analysis will eliminate PCR product contamination problems.

High-throughput MethyLight-based methods allowing the analysis of methylation at hundreds of sites at a time will challenge DNA methylation arrays as a method to analyze methy-lation at multiple sites, particularly because arrays are unlikely to be quantitative over a similar dynamic range. The information gained by multilocus methylation profiling will complement expression profiling as a diagnostic and prognostic tool. Finally, the ultimate methylation analysis method might be based on mass spectrometry that can be done directly from DNA without bisulfite modification.

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