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Multiplex ligation-dependent probe amplification using a completely synthetic probe set.

Rowena F Stern·Roland G Roberts·Kathy Mann·Shu C Yau·Jonathan Berg·Caroline Mackie Ogilvie
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Abstract

The recent development of multiplex ligation-dependent probe amplification (MLPA) has provided an efficient and reliable assay for dosage screening of multiple loci in a single reaction. However, a drawback to this method is the time-consuming process of generating a probe set by cloning in single-stranded bacteriophage vectors. We have developed a synthetic probe set to screen for deletions in a region spanning 18.5 Mb within chromosome 3q. In a pilot study, we tested 15 synthetic probes on 4 control samples and on 2 patients previously found to possess a heterozygous deletion in the region 3q26-q28. These synthetic probes detected deletions at all previously known deleted loci. Furthermore, using synthetic probes, the variability of results within samples was similar to that reported for commercially available M13-derived probes. Our results demonstrate that this novel approach to MLPA provides a generic solution to the difficulties of probe development by cloning; such synthetically generated probes may be used to screen a large number of loci in a single reaction. We conclude that the use of synthetic probes for MLPA is a rapid, robust, and efficient alternative for research (and potentially diagnostic) deletion and duplication screening of multiple genomic loci.

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Chromosome DeletionDNA Mutational AnalysisDNA ProbesGene DosageNucleic Acid Amplification TechniquesJournal ArticleResearch Support, Non-U.S. Gov'tTechnical Report

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Rowena F Stern, Roland G Roberts, Kathy Mann, Shu C Yau, Jonathan Berg, Caroline Mackie Ogilvie. 2004. Multiplex ligation-dependent probe amplification using a completely synthetic probe set.. https://doi.org/10.2144/04373st04

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