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Rapid detection of genomic,,,duplications and deletions using,,,the Agilent 2100 bioanalyzer

d. The DNA is labeled with either radioactive or fluorescent molecules during the PCR step, and the separation has been performed by electrophoresis, either on a polyacrylamide gel or capillary system. We have tried the possibility of analyzing the MLPA and MAPH products using the Agilent 2100 bioanalyzer in combination with the DNA 500 chip. Parameters under investigation included resolution, reproducibility and the accuracy of quantitation.
Methods
MAPH
A total of 17 female DNA samples were examined, of whom 13 were known to be carriers for mutations in the DMD gene and 4 were non-carriers. The MAPH protocol has been described previously3, and is also available on http://www.dmd.nl/DMD_MAPH.html For each hybridization several DMD gene exon probes were chosen, both within and outside the suspected breakpoints. At least 2 control autosomal probes were also included in the probe mix.
MLPA
Full details regarding the MLPA protocol are in reference 2, and on the website http://www.mrc-holland.nl/ The MLPA reaction was performed on male and female DNA, as well as DNA from a cell line 49,XXXXY. Bioanalyzer analysis was performed according to the manufacturer's instructions. The peak patterns were visually compared using the Bio Sizing Data Evaluation program (Agilent Technologies).
Results and discussion

We have previously published work on the analysis of deletions and duplications in the DMD geneusing MAPH3. While this concentrated on the analysis of a large group of samples, it is often necessary or sufficient to screen only one or a small number of samples with a few probes. This is particularly the case when a relative of a patient wishes to be screened for carr
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