Rapid and Inexpensive Screening of Genomic Copy Number Variations Using a Novel Quantitative Fluorescent PCR Method
Figure 1
Experimental design and visualization of microdeletion detection using MQF-PCR. (a) Schematic representation of MQF-PCR primer design. Control region with high sequence similarity to the microdeletion critical region and containing 1–10 bp insertion/deletion (empty box) is used to design MQF-PCR primer pair (arrows) that amplifies two fragments of distinct size. The forward primer is extended at 5′ end using M13-40 universal sequence (black box) to allow cost-efficient M13-40-NED fluorescent labeling and reliable quantification of fluorescent signal intensity, while the reverse primer is extended at 5′ end using the PIG-tail (grey box) to improve adenylation of the 3′ end of the forward strand. (b)–(i) Representative electropherograms showing the change in peak areas corresponding to the syndrome-related chromosomes (black) and the peaks representing the control chromosomes (white) between controls and affected individuals. In all panels, approximately 50% reduction in the peak areas in the affected individuals is observed. Approximate chromosomal locations of the MQF-PCR amplicons are shown.
