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RISS 인기검색어
- 검색키워드
- 저자 : Richard A. Mathies (검색결과 5 건)
- 무료
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Fluorescence energy transfer-labeled primers for high-performance forensic DNA profiling
Yeung, Stephanie H. I.,Seo, Tae Seok,Crouse, Cecelia A.,Greenspoon, Susan A.,Chiesl, Thomas N.,Ban, Jeff D.,Mathies, Richard A. WILEY-VCH Verlag 2008 Electrophoresis Vol.29 No.11
<P>A fluorescence energy transfer (ET) dye-labeled STR typing system (ET 16-plex) is developed for the markers used in the commercial STR typing kit PowerPlex 16, and its performance assessed using a 96-lane microfabricated capillary array electrophoresis (μCAE) system. The ET 16-plex amplicons displayed 1.6–9-fold higher fluorescence intensities compared to those produced using the single-dye (SD)-labeled multiplex kits. The ET multiplex delivered full STR profiles from 62.5 pg of DNA; half the input required for the SD kits while maintaining a similar heterozygote allele balance. This increased sensitivity should improve typing of poor-quality DNA samples by making minor or imbalanced alleles more readily detectable at the low copy number (LCN) threshold. The ET 16-plex also generated complete profiles with only 28 PCR cycles; this capability should improve LCN typing by reducing the amplification time and drop-in allele incidence. To confirm the practical advantages of ET-labeled primers, six previously problematic casework samples were tested and only the ET 16-plex kit was able to capture additional allele data. The successful development and demonstration of ET primers for higher sensitivity STR typing offers a simple solution to improving current commercial multiplex typing capability. The superior spectral properties and universal compatibility with any primer sequence provided by ET cassettes will make future multiplex construction more facile and straightforward. The pairing of ET cassette technology with the μCAE system illustrates not only an enhanced STR typing platform, but a significant step toward a higher-efficiency forensic laboratory enabled by better chemistry and microfluidics.</P>
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Multiplex Mini Y Short Tandem Repeat Haplotyping using Fluorescence Energy Transfer Labeled Primers
서태석,신경진,최종영,Richard A. Mathies 한국바이오칩학회 2008 BioChip Journal Vol.2 No.3
A fluorescence energy transfer (ET) cassette labeled mini Y short tandem repeat (STR) genotyping system is presented. A capillary electrophoretic (CE) microdevice with a cross-injector design is used to determine the fluorescence intensities of ET and single dye-labeled STR amplicons, demonstrating a 2-12 fold higher fluorescence signal of ET cassette labels than the corresponding single dye-labeled ones. Eleven extended haplotype mini Y-STRs using ET cassette labeled primers are constructed, and sensitivity and mixed sample studies are performed. Due to the improved spectroscopic properties of ET labels, multiplex mini Y-STR typing is successful with as low as 30 pg of genomic male DNA, and in the high background of female DNA. These results indicate the practical advantage of ET cassette labels for low copy number and poor-quality DNA STR genotyping to be applied for criminal investigations, paternity testing, and evolutionary studies.
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Microfluidic Linear Hydrogel Array for Multiplexed Single Nucleotide Polymorphism (SNP) Detection
Jung, Yun Kyung,Kim, Jungkyu,Mathies, Richard A. American Chemical Society 2015 ANALYTICAL CHEMISTRY - Vol.87 No.6
<P>A PDMS-based microfluidic linear hydrogel array is developed for multiplexed single nucleotide polymorphism (SNP) detection. A sequence of three-dimensional (3D) hydrogel plugs containing the desired DNA probes is prepared by UV polymerization within a PDMS microchannel system. The fluorescently labeled target DNA is then electrophoresed through the sequence of hydrogel plugs for hybridization. Continued electrophoresis provides an electrophoretic wash that removes nonspecific binders. The capture gel array is imaged after washing at various temperatures (temperature gradient electrophoresis) to further distinguish perfect matches from mismatches. The ability of this microdevice to perform multiplex SNP genotyping is demonstrated by analyzing a mixture of model <I>E. coli</I> bacterial targets. This microfluidic hydrogel array is ∼1000 times more sensitive than planar microarrays due to the 3D gel capture, the hybridization time is much shorter due to electrophoretic control of the transport properties, and the stringent wash with temperature gradient electrophoresis enables analysis of single nucleotide mismatches with high specificity.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/ancham/2015/ancham.2015.87.issue-6/ac5048696/production/images/medium/ac-2014-048696_0005.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/ac5048696'>ACS Electronic Supporting Info</A></P>
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