Capillary electrophoresis combining three-step multiplex polymerase chain reactions for diagnosing α-thalassemia
作者： Yen-Ling Chen
关键词:
摘要: Capillary electrophoresis (CE) is the most useful tool for DNA separation because of its high resolution. In this study, different kinds polymers were used to evaluate efficiency by analyzing a 200-bp ladder. Under optimized CE conditions, was performed DB-17 capillary. The running buffer 1× TBE containing 0.6% w/v poly(ethylene oxide) (PEO) (Mw: 8,000,000) and 1 μM YO-PRO-1; applied voltage -10 kV (detector at anode side) temperature 25°C. these optimal 15 fragments with sizes ranging from 0.2 3.0 kb resolved within 11.5 min RSD migration time less than 0.55% (n=3). This method, combined three-step multiplex PCR, detect five α-thalassemia deletions, including -α(3.7) , -α(4.2) - -(SEA) -(FIL) -(THAI) . A total 21 patients diagnosed analyzed using developed method all results agreed those already obtained gel electrophoresis.
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