کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6553771 | 160893 | 2015 | 18 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
High sensitivity multiplex short tandem repeat loci analyses with massively parallel sequencing
ترجمه فارسی عنوان
تجزیه و تحلیل تکرار چند ضلعی با حساسیت بالا چند ضلعی با توالی بسیار موثر
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
چکیده انگلیسی
STR typing in forensic genetics has been performed traditionally using capillary electrophoresis (CE). However, CE-based method has some limitations: a small number of STR loci can be used; stutter products, dye artifacts and low level alleles. Massively parallel sequencing (MPS) has been considered a viable technology in recent years allowing high-throughput coverage at a relatively affordable price. Some of the CE-based limitations may be overcome with the application of MPS. In this study, a prototype multiplex STR System (Promega) was amplified and prepared using the TruSeq DNA LT Sample Preparation Kit (Illumina) in 24 samples. Results showed that the MinElute PCR Purification Kit (Qiagen) was a better size selection method compared with recommended diluted bead mixtures. The library input sensitivity study showed that a wide range of amplicon product (6-200Â ng) could be used for library preparation without apparent differences in the STR profile. PCR sensitivity study indicated that 62Â pg may be minimum input amount for generating complete profiles. Reliability study results on 24 different individuals showed that high depth of coverage (DoC) and balanced heterozygote allele coverage ratios (ACRs) could be obtained with 250Â pg of input DNA, and 62Â pg could generate complete or nearly complete profiles. These studies indicate that this STR multiplex system and the Illumina MiSeq can generate reliable STR profiles at a sensitivity level that competes with current widely used CE-based method.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Forensic Science International: Genetics - Volume 16, May 2015, Pages 38-47
Journal: Forensic Science International: Genetics - Volume 16, May 2015, Pages 38-47
نویسندگان
Xiangpei Zeng, Jonathan L. King, Monika Stoljarova, David H. Warshauer, Bobby L. LaRue, Antti Sajantila, Jaynish Patel, Douglas R. Storts, Bruce Budowle,