A simplified procedure for developing multiplex PCRs.
TLDR
The use of tagged primers provides a method for primer design that eliminates the multiple optimization steps involved in developing multiplex PCR.Abstract:
We have developed a simplified method for multiplex PCR based on the use of chimeric primers. Each primer contains a 3' region complementary to sequence-specific recognition sites and a 5' region made up of an unrelated 20-nucleotide sequence. Identical reaction conditions, cycling times, and annealing temperatures have been established for any PCR primer pair comprising the chimeric motif. Under these conditions, efficient multiplex amplification is achieved easily and reproducibly by simple adjustment of the individual primer concentrations. No additional modification of either the reaction components or annealing temperatures is required. The use of tagged primers provides a method for primer design that eliminates the multiple optimization steps involved in developing multiplex PCR.read more
Citations
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Journal ArticleDOI
Point-of-care diagnostics for global health.
TL;DR: The context in which the diagnostics must operate, some of the appropriate diagnostic technologies already in distribution, and some emerging technologies that promise to address this challenge are reviewed.
Journal ArticleDOI
Multiplex PCR: Optimization and Application in Diagnostic Virology
TL;DR: The principles, optimization, and application of multiplex PCR for the detection of viruses of clinical and epidemiological importance are reviewed.
Patent
Droplet-based assay system
Billy W. Colston,Benjamin J. Hindson,Kevin D. Ness,Donald A. Masquelier,Fred P. Milanovich,Douglas N. Modlin,Vincent J. Riot,Samuel Burd,Anthony J. Makarewicz,Phillip Belgrader +9 more
TL;DR: In this article, the authors present a system for separating sample components by partitioning them into droplets or other partitions, amplifying or reacting the components within the droplets, detecting the amplified components, or characteristics thereof, and/or analyzing the resulting data, among others.
Journal ArticleDOI
Molecular Typing of Enteroviruses: Current Status and Future Requirements
Peter Muir,Ulrike Kämmerer,Klaus Korn,Mick N. Mulders,Tuija Pöyry,Benedikt Weissbrich,Reinhard Kandolf,Graham M. Cleator,Anton M. van Loon +8 more
TL;DR: The case for developing a molecular typing system is argued, the genetic basis of such a system is discussed, the literature describing attempts to identify or classify enteroviruses by molecular methods is reviewed, and ways in which the goal of molecular typing may be realized are suggested.
Journal ArticleDOI
A second-generation genetic linkage map of the domestic dog, Canis familiaris.
Mark W. Neff,Karl W. Broman,Cathryn S. Mellersh,Kunal Ray,Gregory M. Acland,Gustavo D. Aguirre,Janet S. Ziegle,Elaine A. Ostrander,Jasper Rine +8 more
TL;DR: A canine linkage map with the number of mapped loci expanded to 276 and 10-cM coverage extended to 75-90% of the genome is reported, with fifteen markers anchored well-described genes on the map, thereby serving as landmarks for comparative mapping in dogs.
References
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Journal ArticleDOI
DNA sequencing with chain-terminating inhibitors
TL;DR: A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.
Book ChapterDOI
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Allan M. Maxam,Walter Gilbert +1 more
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Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms
TL;DR: The mobility shift analysis of single-stranded DNAs on neutral polyacrylamide gel electrophoresis to detect DNA polymorphisms was developed and SSCPs were found to be allelic variants of true Mendelian traits, and therefore they should be useful genetic markers.
Journal ArticleDOI
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Batsheva Kerem,Johanna M. Rommens,Janet A. Buchanan,Danuta Markiewicz,Tara K. Cox,Aravinda Chakravarti,Manuel Buchwald,Lap-Chee Tsui +7 more
TL;DR: Extended haplotype data based on DNA markers closely linked to the putative disease gene locus suggest that the remainder of the cystic fibrosis mutant gene pool consists of multiple, different mutations.
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