1. Array Comparative Genomic Hybridization (Array CGH) for Detection of Genomic Copy Number Variants
- Author
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Michael Coldwell, Caroline Mackie Ogilvie, Susan Bint, and Joo Wook Ahn
- Subjects
Comparative Genomic Hybridization ,DNA Copy Number Variations ,General Immunology and Microbiology ,Genome, Human ,Oligonucleotide ,Chemistry ,General Chemical Engineering ,General Neuroscience ,Hybridization probe ,Copy number analysis ,High-Throughput Nucleotide Sequencing ,DNA ,Genome ,Molecular biology ,General Biochemistry, Genetics and Molecular Biology ,Karyotyping ,Humans ,Human genome ,Copy-number variation ,DNA Probes ,Molecular Biology ,Virtual karyotype ,Comparative genomic hybridization - Abstract
Array CGH for the detection of genomic copy number variants has replaced G-banded karyotype analysis. This paper describes the technology and its application in a clinical diagnostic service laboratory. DNA extracted from a patient’s sample (blood, saliva or other tissue types) is labeled with a fluorochrome (either cyanine 5 or cyanine 3). A reference DNA sample is labeled with the opposite fluorochrome. There follows a cleanup step to remove unincorporated nucleotides before the labeled DNAs are mixed and resuspended in a hybridization buffer and applied to an array comprising ~60,000 oligonucleotide probes from loci across the genome, with high probe density in clinically important areas. Following hybridization, the arrays are washed, then scanned and the resulting images are analyzed to measure the red and green fluorescence for each probe. Software is used to assess the quality of each probe measurement, calculate the ratio of red to green fluorescence and detect potential copy number variants.
- Published
- 2015