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DNA Microarrays02:34

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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Technical Demonstration of Whole Genome Array Comparative Genomic Hybridization
16:37

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Published on: August 5, 2008

Comparative genomic hybridization on BAC arrays.

Bradley P Coe1, William W Lockwood, Raj Chari

  • 1British Columbia Cancer Research Centre, BC, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|June 3, 2009
PubMed
Summary

Array comparative genomic hybridization (aCGH) using bacterial artificial chromosomes (BACs) enables precise detection of genomic copy number alterations. This method aids in diagnosing genetic disorders and cancer by identifying crucial gene dosage regions.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Medical Genetics

Background:

  • Genomic DNA alterations are central to constitutional disorders and cancer.
  • Identifying gene dosage regions is critical for understanding disease phenotypes and developing diagnostics.
  • Array comparative genomic hybridization (aCGH) has emerged as a powerful tool for genomic analysis.

Purpose of the Study:

  • To detail the technical procedure for performing copy number analysis using BAC aCGH.
  • To highlight the utility of BAC aCGH in discovering and fine-mapping genomic alterations.
  • To emphasize the application of BAC aCGH in identifying target genes for diagnostics and therapeutics.

Main Methods:

  • DNA samples are labeled with fluorescent dyes via random priming.
  • Labeled DNA probes are co-hybridized to arrays of bacterial artificial chromosome (BAC) clones.
  • BAC clone arrays offer high probe-binding capacity (approx. 150 kbp) with resolutions of ~50 kbp or ~1 Mbp.

Main Results:

  • BAC aCGH facilitates the discovery and fine mapping of novel genomic alterations.
  • The method allows for rapid identification of target genes associated with copy number variations.
  • BAC aCGH requires significantly lower sample input compared to oligonucleotide platforms.

Conclusions:

  • BAC aCGH is an effective technique for comprehensive genome-wide copy number analysis.
  • This method is valuable for dissecting complex genetic disorders and identifying cancer-related genomic changes.
  • The detailed protocol described enables researchers to implement BAC aCGH for diagnostic and research purposes.