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Genotyping of exons 1 to 20 in Duchenne muscular dystrophy by universal multiplex PCR and short‐end capillary electrophoresis

Author:
Syu, Jing‐Rou, Wang, Chun‐Chi, Jong, Yuh‐Jyh, Wu, Shou‐Mei
Source:
Electrophoresis 2014 v.35 no.23 pp. 3387-3394
ISSN:
0173-0835
Subject:
capillary electrophoresis, exons, fluorescence, genes, genotyping, males, muscular dystrophy, patients, polymerase chain reaction, polymers
Abstract:
One rapid CE method was established to diagnose Duchenne muscular dystrophy (DMD). DMD is a severe recessive inherited disorder frequently caused by gene deletions. Among them, exons 1–20 account for nearly 30% of occurrences. In this study, the universal multiplex PCR was used to enhance the fluorescently labeling efficiency, which was performed only by one universal fluorescent primer. After PCR, a short‐end injection CE (short‐end CE) speeded up the genotyping of the DMD gene. This method involved no extra purification, and was completed within 9 min. The CE conditions contained a polymer solution of 1.5% hydroxylethylcellulose in 1× TBE buffer at 6 kV for separation. This method was applied to test six DMD patients and one healthy male person. The results showed good agreement with those of multiplex ligation‐dependent probe amplification. This method can be applied for clinical diagnosis of DMD disease. Accurate diagnosis of the DMD gene is the best way to prevent the disease.
Agid:
1323135