Journal ArticleDOI
Fast multiplexed polymerase chain reaction for conventional and microfluidic short tandem repeat analysis.
TLDR
The fast multiplex PCR approach has the potential to reduce process time and cost for STR analysis and enables development of a fully integrated microfluidic forensic DNA analysis system.Abstract:
The time required for short tandem repeat (STR) amplification is determined by the temperature ramp rates of the thermal cycler,the components of the reaction mix, and the properties of the reaction vessel. Multiplex amplifications in microfluidic biochip-based and conventionaltube-based thermal cyclers have been demonstrated in 17.3 and 19 min, respectively. Optimized 28-cycle amplification protocols generated alleleswith signal strengths above calling thresholds, heterozygous peak height ratios of greater than 0.65, and incomplete nontemplate nucleotide additionand stutter of less than 15%. Full CODIS-compatible profiles were generated using the Profiler Plus ID, COfiler and Identifiler primer sets. PCR per-formance over a wide range of DNA template levels from 0.006 to 4 ng was characterized by separation and detection on a microfluidic electropho-resis system, Genebench-FXTM. The fast multiplex PCR approach has the potential to reduce process time and cost for STR analysis and enablesdevelopment of a fully integrated microfluidic forensic DNA analysis systemread more
Citations
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Journal ArticleDOI
Point-of-care testing (POCT) diagnostic systems using microfluidic lab-on-a-chip technologies
TL;DR: Categorize the applications of the POCT diagnostic systems according to the analyte types such as proteins, cells, nucleic acids, and metabolites, and advantages and disadvantages of POCT systems are reviewed and discussed along with future prospects.
Journal ArticleDOI
Integrated DNA purification, PCR, sample cleanup, and capillary electrophoresis microchip for forensic human identification
TL;DR: A fully integrated microdevice and process for forensic short tandem repeat (STR) analysis has been developed that includes sequence-specific DNA template purification, polymerase chain reaction (PCR), post-PCR cleanup and inline injection, and capillary electrophoresis (CE).
Multiplex PCR: Critical Parameters and Step-by-Step Protocol
TL;DR: Based on the experience, a protocol for developing a multiplex PCR assay is proposed and ways to overcome commonly encountered problems are suggested.
Patent
Nucleic acid purification
Richard F. Selden,Eugene Tan +1 more
TL;DR: In this article, a self-contained apparatus for isolating nucleic acid, cell lysates and cell suspensions from unprocessed samples apparatus, to be used with an instrument, comprises at least one input, and a drive mechanism on said instrument for driving said liquid purification reagent, through a microfluidic element and a purification matrix, wherein the only inputs to said apparatus are via said chamber and said drive mechanism.
Journal ArticleDOI
A protocol for direct and rapid multiplex PCR amplification on forensically relevant samples
TL;DR: Direct and rapid PCR is incorporated in a "DNA-6h" service that can assist police investigations by rapidly deriving DNA information from trace evidence left by a perpetrator, searching the STR profile against a DNA database and reporting the outcomes to police or prosecution.
References
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Randall Keichi Saiki,Stephen J. Scharf,Fred A. Faloona,Kary B. Mullis,Glenn Thomas Horn,Henry A. Erlich,Norman Arnheim +6 more
TL;DR: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia, using primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase of target DNA copies.
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Chemical Amplification: Continuous-Flow PCR on a Chip
TL;DR: A micromachined chemical amplifier was successfully used to perform the polymerase chain reaction (PCR) in continuous flow at high speed.
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TL;DR: This introductory guide covers the basic methodology, research applications, and medical applications of PCR technology, emphasizing the practical aspects of the technique.
Journal ArticleDOI
Modulation of non-templated nucleotide addition by Taq DNA polymerase: primer modifications that facilitate genotyping.
TL;DR: It was clear, however, that features of the template in addition to the 3' terminal base also affect the fraction of product adenylated, and it proved difficult to identify a single sequence capable of protecting the products of all markers from non-templated addition of nucleotide.
Journal Article
DNA typing and genetic mapping with trimeric and tetrameric tandem repeats.
TL;DR: A method enabling rapid localization of STRs and determination of their flanking DNA sequences was developed, thus simplifying the identification of polymorphic STR loci.