calculated. The ranges are such that the actual error rate is likely to be lower than the predicted rate of 1 %. It should also be possible to detect mosaic cases i.e. when the mutation is not present in all of the cells and is therefore masked by the presence of normal cells. In these cases the p-value may fall into the retest range. In such cases alternative methods such as FISH could be employed to confirm this.
We also looked at testing MLPA reactions on the DNA 500 chip. Initial tests were performed with a test kit provided by MRC Holland (Nederland). The resulting peaks can be analyzed in exactly the same way as those obtained from a MAPH experiment. An example of the results can be seen in figure 3. The difference in the number of X chromosomes can be easily seen as differences in peak height of the appropriate probes. In this regard the software provided (BioSizing Data Evaluation) makes it very easy to overlay different traces and make direct visual comparisons.
Conclusion
These experiments show that the Lab-on-a-chip combined with MAPH /MLPA is a quick and simple way to detect copy number changes in genomic DNA. Particular strengths are the speed and relative automation of analysis. The resolution is sufficient to separate up to 15 probes accurately on the DNA 500 chip, more than sufficient when examining only one particular area of the genome. We consider this technique to be an attractive alternative to FISH in a diagnostic setting, particularly for mutations that cannot be readily detected in such a manner.



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