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Microarray MAPH: accurate array-based detection of relative copy number in genomic DNA.
Brian Gibbons1, Parikkhit Datta, Ying Wu
1Institute of Genetics, University of Nottingham, QMC, Nottingham NG7 2UH, UK. drgibbo@ntlworld.com
BMC Genomics
|July 4, 2006
Summary
This study introduces a novel microarray-based method for Multiplex Amplifiable Probe Hybridisation (MAPH) to accurately measure gene copy number. The new approach enhances throughput and simplifies analysis for genetic disorders like Charcot-Marie-Tooth disease type 1A.
Area of Science:
- Genetics
- Molecular Biology
- Bioinformatics
Background:
- Current copy number measurement methods lack convenience, throughput, economy, accuracy, and resolution.
- Multiplex Amplifiable Probe Hybridisation (MAPH) offers potential but requires throughput improvement.
Purpose of the Study:
- To enhance Multiplex Amplifiable Probe Hybridisation (MAPH) throughput using a 3-Dimensional, Flow-Through Microarray Platform.
- To replace electrophoretic analysis with photometric analysis for improved efficiency.
Main Methods:
- Developed a modified MAPH technique utilizing a microarray platform.
- Employed a dual-label approach with Cy3 and Cy5 probes for comparative analysis.
- Utilized sequence-based discrimination of amplified probes instead of fragment length.
Main Results:
- Microarray MAPH measurement of PMP22 gene dosage showed strong correlation with capillary MAPH.
- Accurate copy number reporting was achieved in DNA analyses from 38 individuals, including 12 with Charcot-Marie-Tooth disease type 1A (CMT1A).
Conclusions:
- Microarray-based MAPH offers comparable accuracy to electrophoretic methods.
- This technology can fully exploit MAPH's potential multiplicity for complex genetic analyses.
- The method simplifies copy number assays for genes with multiple exons or dispersed genomic locations.