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Chemically Modified Primers for Improved Multiplexed PCR
Elena Hidalgo Ashrafi, Tony Le, Alexandre Lebedev, Richard Hogrefe, Victor Timoshchuk, Sabrina Shore, Inna Koukhareva and Natasha Paul,
TriLink BioTechnologies,
Multiplex PCR is an advantageous technique used in PCR applications to amplify multiple targets in a single reaction. As useful as it is, this technique presents a new set of challenges that further complicates PCR setup. For example, reactions are more prone to off-target amplifications such as mis-priming and primer dimer due to the increased number of primer pairs. Furthermore, preferential amplification of certain targets leads to an unequal distribution of amplicon products, making quantifi
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Hot Start dNTPs - A Novel Tool for Controlled Nucleotide Incorporation in PCR
Tony Le, Elena Hidalgo Ashrafi, Sabrina Shore, Victor Timoshchuk, Natasha Paul, Richard Hogrefe, Inna Koukhareva, Alexandre Lebedev,
TriLink BioTechnologies,
PCR is a widely used scientific tool employed by a variety of applications. Various Hot Start technologies have already been developed using modified PCR components to increase specificity of a reaction. Recently developed CleanAmpTM dNTPs are modified nucleoside triphosphates with a thermolabile 3’-tetrahydrofuranyl protecting group that is released at higher temperatures. These modified dNTPs prevent low temperature primer extension, which can often be a significant problem in PCR. At higher t
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Evaluation of microfluidic digital PCR for the detection of cancer biomarkers
Rebecca Sanders, Claire Bushell, Carole Foy, Daniel J. Scott,
LGC,
dPCR is achieved by sample partitioning prior to PCR amplification such that each reaction chamber contains one copy or less of target DNA. This dilution becomes the limiting factor and an accurate target molecule count is achievable. This study evaluates dPCR’s quantitative capabilities and investigates parameters influencing copy number quantification, using the Fluidigm Biomark instrument. Biomark technology combines dPCR theory with a microfluidics platform.
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A novel multiplex qPCR for the simultaneous detection of Salmonella spp., E. coli O157:H7 and Listeria monocytogenes from raw Atlantic Salmon fillets
Sonja S. Nitecki, Brian F. Carney, John W. Slater, Wolfram M. Bruck,
CAMBio, Letterkenny Institute of Technology,
A triplex qPCR assay was developed to screen for three major food-borne pathogens in fish. The assay is based on simultaneous qPCR amplification of specific target genomes using fluorophore labelled locked nucleic acid (LNA) TaqMan probes. The detection limit of the qPCR is 1 colony forming unit (cfu) per 1ml and the detection limit of the assay with 24h general enrichment is 1 cfu per 25g of fish meat.
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SYBR® Green I as a biomarker for schizophrenia
Davood Zaeifi, Mehdi Rahmati, Vahab Piranfar,
Islamic Azad University of Tonekabon,
The purpose of this study were to examine if the mRNA of the peripheral dopamine receptor is changed in schizophrenia patients. 50 Naive patients were enrolled. After extracting RNA from white blood cells, cDNA is synthesized. After doing the quantitative Real-time PCR, expression of D3 receptors in healthy persons and patients were compared. Results of this examinations reveals that expression of D3 receptor is increased in compared with controls sample.
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Evaluation of a Novel Approach for the Measurement of RNA Quality
Timothy Wilkes; Alison Devonshire; Carole Foy,
LGC Limited,
The potential medical applications of microarrays have generated much interest, within the biomedical community. Numerous potential sources of variation have raised concerns regarding assay consistency and data quality. Previous studies have highlighted RNA integrity as having a major effect on data quality. We describe here a comparison of the performance characteristics of the Agilent (Bioanalyser) and Lab901 (Screen Tape System) platforms, and their capacity to determine sample RNA integrity.
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