DNA Sequence Fragment Containing C to A Mutation as a Convenient Mutation Standard for DHPLC Analysis
Publish place: Iranian Journal of Basic Medical Sciences، Vol: 16، Issue: 8
Publish Year: 1392
نوع سند: مقاله ژورنالی
زبان: English
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شناسه ملی سند علمی:
JR_IJBMS-16-8_011
تاریخ نمایه سازی: 4 آبان 1400
Abstract:
Objective(s): Denaturing high performance liquid chromatography (DHPLC) is a high throughput approach for screening DNA sequence variations. To assess oven calibration, cartridge performance, buffer composition and stability, the WAVE Low and High Range Mutation Standards are employed to ensure reproducibility and accuracy of the chromatographic analysis. The purpose of this study was to provide a cost-effective homemade mutation standard for DHPLC analysis. Materials and Methods: DHPLC was performed to evaluate different elution temperatures of a ۳۷۴ bp DNA fragment with C>A mutation at position of ۵۹ to achieve a peak profile similar to the Low Mutation Standard. In order to verify the reproducibility of the homemade mutation standard using DHPLC, ۱۵ different experiments were performed to compare the homemade mutation standard, the WAVE Low Range Mutation Standard with a positive DNA control sample. Results:We identified a comparable elution temperature and a peak profile with the WAVE Low Range Mutation Standard. Conclusion: This study confirmed the reproducibility of the peak profile of our homemade mutation standard compared to the Low Mutation Standard using DHPLC analysis.
Keywords:
ATP۷B DHPLC Low range mutation stan- dar
Authors
Hassan Dastsooz
۱Department of Medical Genetics, Shiraz University of Medical Sciences, Shiraz, Iran
Nazanin Vahedi
۱Department of Medical Genetics, Shiraz University of Medical Sciences, Shiraz, Iran
Majid Fardaei
۱Department of Medical Genetics, Shiraz University of Medical Sciences, Shiraz, Iran
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