کد مقاله کد نشریه سال انتشار مقاله انگلیسی ترجمه فارسی نسخه تمام متن
5519709 1544412 2017 3 صفحه PDF سفارش دهید دانلود کنید
عنوان انگلیسی مقاله ISI
Random shearing as an alternative to digestion for mitochondrial DNA processing in droplet digital PCR
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوفیزیک
پیش نمایش صفحه اول مقاله
Random shearing as an alternative to digestion for mitochondrial DNA processing in droplet digital PCR
چکیده انگلیسی


- Droplet digital PCR (ddPCR) is a quantitative assay that requires DNA fragmentation to maximize reaction efficiency.
- We measured the proportion of mtDNA carrying the “common deletion,” a rare event associated with aging and neurodegeneration.
- We compared alternative fragmentation methods (sonication and QIAshredder columns) against enzymatic digestion.
- QIAshredder showed the highest sensitivity when compared to sonication, followed by digestion.
- QIAshredder and sonication are fragmentation methods that maximize ddPCR quantification for the detection of rare events.

Droplet digital PCR (ddPCR) is a quantitative assay that requires DNA fragmentation to maximize reaction efficiency. Here, we measured the proportion of mitochondrial DNA (mtDNA) carrying the “common deletion,” a rare event, to compare quantification sensitivities between alternative DNA fragmentation methods (sonication and QIAshredder spin columns) against enzymatic digestion (traditionally used). QIAshredder showed the highest sensitivity when compared to sonication, followed by digestion. Also, both sonication and QIAshredder fragmentation had shorter processing times than enzymatic digestion; therefore, QIAshredder fragmentation and sonication are alternative DNA processing methods that maximize ddPCR quantification for the detection of rare events.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Mitochondrion - Volume 32, January 2017, Pages 16-18
نویسندگان
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