|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
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
|Evaluation of microfluidic digital PCR for the detection of cancer biomarkers|
Rebecca Sanders, Claire Bushell, Carole Foy, Daniel J. Scott
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.
|Inhibition of DNA methylation does not overcome docetaxel resistance in human breast cancer cells|
Kastl L, Brown I, Schofield A
DNA mehtylation can lead to chemotherapy resistance in cancer. The aim of this study was to investigate the role of DNA methylation in docetaxel-resistant human breast cancer cells. Whereas the DNA methylation machinery is altered in docetaxel-resistant human breast cancer cells compared to docetaxel-sensitive human breast cancer cells, resistance to docetaxel could not be reversed using the DNA methylation inhibitor decitabine.
|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
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.
|Insulin Decreases Transcription of Three Proteins Associated with Lung Surfactant|
Rucka, Z [a], Vanhara, P [b], Tesarova, L [a], Potesilova, M [a], Stejskal, S [a], Dolezel, J [c], Koutna, I [a]
Our project is partially focused on insulin affecting four proteins associated with lung surfactant. Small and hydrophobic SP-B and S-PC help to spread and stabilize the surfactant layer while large and hydrophilic SP-A and SP-D participate in immune responses. A549 and H441 cell lines were used. It was observed using RealTime PCR that higher doses of insulin lead to decreased transcription of SP-B, SP-C, SP-D, and both isoforms of SP-A.
|Tapping into the mechanical links of expansins in leaf morphogenesis|
Expansins are conserved family of non-enzymatic proteins involved in cell wall-loosening accounted for widespread role in plant development. My research focuses on functional analysis of expansins during leaf morphogenesis. Atomic force microscopy was applied to characterise cell wall mechanical properties in an attempt for in vivo quantification of expansin activity. It’s showed how that there is a distinct difference in cell wall extensibility between leaf abaxial and adaxial side.
|SYBR® Green I as a biomarker for schizophrenia|
Davood Zaeifi, Mehdi Rahmati, Vahab Piranfar
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.
|Evaluation of a Novel Approach for the Measurement of RNA Quality|
Timothy Wilkes; Alison Devonshire; Carole Foy
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.
|Microfluidic PCR device for diagnostic pathogen detection|
Johannes R. Peham, Hannes Steiner, Walter Grienauer, Rudolf Heer, Michael J. Vellekoop, Christa Nöhammer, Herbert Wiesinger
In this work a microfluidic cyclic flow PCR device is presented, which is capable of replicating the bacterial genomic DNA sequence of the 16S ribosomal RNA. The standard laboratory processing time of 3 h could be decreased to 60 min with the microfluidic reactor without loosing PCR efficiency. Integrating an optical fluorescence detector for dsDNA measurement would evolve this device into a micro total analysis system.