کد مقاله کد نشریه سال انتشار مقاله انگلیسی ترجمه فارسی نسخه تمام متن
98711 160877 2016 15 صفحه PDF سفارش دهید دانلود رایگان
عنوان انگلیسی مقاله ISI
Validation of alternative capillary electrophoresis detection of STRs using POP-6 polymer and a 22 cm array on a 3130xl genetic analyzer
ترجمه فارسی عنوان
اعتبارسنجی از تشخیص الکتروفورز STRs مویرگی جایگزین با استفاده از پلیمر POP-6 و یک آرایه 22 سانتی متر در یک تجزیه و تحلیل ژنتیکی 3130xl
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تولید محتوا
با 10 درصد تخفیف ویژه دانشیاری
کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی ژنتیک
چکیده انگلیسی


• A detection method is presented that utilizes POP-6 polymer and 22 cm array.
• Detection time is reduced by ∼40% and sample throughput is increased ∼67%.
• Full allele concordance is achieved without panel and bin modifications.
• Resolution is slightly improved for smaller sized alleles (<200 b).
• One base resolution is achievable for allele sizes of up to 300 b.

The goal of this project was to reduce capillary electrophoresis detection time on a 3130xl Genetic Analyzer for amplification product obtained from 4-dye and 5-dye STR amplification kits while still generating high quality STR profiles. This was accomplished by utilizing a more viscous polymer (POP-6™) and a shorter array (22 cm) than that which are typically used (POP-4® polymer and a 36 cm array) for human identification purposes. Spatial calibration and detection run modules were modified in response to the use of this polymer/array combination and to reduce detection time. Alternative detection resulted in 24–28 min run times, as compared to ∼45 min using traditional POP-4®/36 cm detection methods. POP-6™/22 cm detection run modules were validated for use with 4-dye Promega STR kits (e.g., PowerPlex® 16 and PowerPlex® 16HS) and 5-dye Life Technologies kits (e.g., Identifiler® and Identifiler® Plus). Three hundred ninety-five samples, controls and allelic ladders were used for the validation studies, which consisted of a comparison of alternative POP-6™/22 cm detection to traditional POP‐4®/36 cm (including reproducibility/concordance of allele calls, resolution, ILS sizing quality, peak height and pass rates), a sizing study (precision and accuracy) and a sensitivity study to obtain a usable range of injection times. Compared to traditional POP-4®/36 cm detection, alternative detection resulted in 100% reproducible and concordant alleles, the ability to achieve one base resolution, slightly reduced ILS sizing quality, slightly reduced peak height and statistically similar pass rates (α = 0.05). It should be noted that alternative detection offered improved resolution over that of traditional for amplicons less than ∼200 b, but had reduced resolution for products greater than ∼200 b. Additionally, alternative detection yielded acceptable precision and accuracy of sizing using Life Technologies criteria (<0.15 standard deviation of allele sizing and ±0.5 b sizing differences for the same allele) and usable injection parameters of 2 kV 4–15 s (compared to 3 kV 10 s for traditional). The run modules developed and validated for 4-dye and 5-dye STR kits using POP‐6™ polymer on a 22 cm array offer a tremendous reduction in detection time (∼40%) while still generating high quality STR profiles.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Forensic Science International: Genetics - Volume 22, May 2016, Pages 113–127
نویسندگان
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