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The DRIgest III (GE Healthcare) molecular weight marker (MW) was used for this experiment, the 4.36, 2.32 and 2.03 kb fragments are indicated by Although the precise localization of the deletion breakpoints that extended over the 5′ and/or 3′ gene boundaries has not been determined, gene dosage assays were performed to estimate the lengths of these sequence changes.Twenty-nine regions were used for copy number analyses from 7.6 MB upstream to 1.5 MB downstream of the ].Included in this study were 87 patients from well-characterized familial adenomatous polyposis kindreds.The majority of cases presented the clinical symptoms of classical FAP, while 21 patients were diagnosed with ], reliable family history information gathered through past clinical records was available only for a minority of the patients/families involved.

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FAP1 is caused by germline mutations of the adenomatous polyposis coli (APC) gene, encoding a multifunctional gatekeeper tumor suppressor protein expressed in a wide variety of tissues.The entire coding region and splice junctions of the gene were amplified by PCR (primer sequences are available upon request).Mutation screening was performed using direct bidirectional sequencing on an ABI 3130 Genetic Analyzer (Life Technologies).At a later stage of our experiments, a different approach was preferred, and copy number determination was done using the robust dosage PCR (RD-PCR) methodology, in which the target locus and an internal control with known copy number are co-amplified [].Briefly, PCR was performed in a duplex or triplex design containing primers for the target locus/loci and for an endogenous control with two copies.

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