Fast detection of deletion breakpoints using quantitative PCR
Loading...
Date
2016
Authors
Abildinova, Gulshara
Abdrakhmanova, Zhanara
Tuchinsky, Helena
Nesher, Elimelech
Pinhasov, Albert
Raskin, Leon
Journal Title
Journal ISSN
Volume Title
Publisher
Genetics and Molecular Biology
Abstract
The routine detection of large and medium copy number variants (CNVs) is well established. Hemizygotic deletions or duplications in the large Duchenne muscular dystrophyDMD gene responsible for Duchenne and Becker muscular dystrophies are routinely identified using multiple ligation probe amplification and array-based comparative
genomic hybridization. These methods only map deleted or duplicated exons, without providing the exact location of breakpoints. Commonly used methods for the detection of CNV breakpoints include long-range PCR and primer walking, their success being limited by the deletion size, GC content and presence of DNA repeats. Here, we present a strategy for detecting the breakpoints of medium and large CNVs regardless of their size. The hemizygous deletion of exons 45-50 in the DMD gene and the large autosomal heterozygous PARK2 deletion were used to demonstrate the workflow that relies on real-time quantitative PCR to narrow down the deletion region and Sanger sequencing for
breakpoint confirmation. The strategy is fast, reliable and cost-efficient, making it amenable to widespread use in genetic laboratories.
Description
Keywords
deletion boundaries, deletion breakpoints, DMD gene, Duchenne and Becker muscular dystrophies, hemizygous deletions, heterozygous deletions
Citation
Abildinova, Gulshara, Abdrakhmanova, Zhanara, Tuchinsky, Helena, Nesher, Elimelech, Pinhasov, Albert , Raskin, Leon. (2016) Fast detection of deletion breakpoints using quantitative PCR. Genetics and Molecular Biology. 39, 3, 365-369 pages