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Proteomic non-small cell lung carcinoma biomarker screening in bronchoalveolar lavage fluid
Tonio Oumeraci 1, Bernd Schmidt 3, Thomas Wolf 1,4, Marc Zapatka 4, Andreas Pich 2, Benedikt Brors 4, Roland Eils 4,5, Brigitte Schlegelberger 1, Nils von Neuhoff 1. 1 Institute of Cell and Molecular Pathology - 2 Department of Toxicology, Hannover Medical School MHH 3 Respiratory Medicine Department of Internal Medicine I, University Hospital Halle, Halle (Saale)

Using a standardized method to acquire MALDI-TOF proteome profile spectra of bronchoalveolar lavage fluid (BALF), we have shown the upregulation of histatin 3 and calgranulin C in a small pilot cohort of NSCLC patients. This pilot study serves to demonstrate that it is feasible to screen a larger NSCLC patient cohort for BALF proteome level biomarkers in a clinical setting.

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

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

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.

Software for Genomic/Epigenomic Research
A.S.Tanas, V.V.Shkarupo, E.B.Kuznetsova, D.V.Zaletayev, V.V. Strelnikov. Research Centre for Medical Genetics; Moscow Medical Academy

We present two computer programs: 1) AIMS in silico - a suggestion tool for Amplification of Intermethylated Sites experimental design and results analysis; and 2) PeakPick – a tool to view & analyse capillary electrophoregrams Our computer software is intended to standardize AIMS applications and to turn it into one of the principal approaches towards cancer methylomes studies as well as towards diagnostics in oncology, including early screening.

HIV POC testing by ssDNA coupled with NALF
Natasha Gous, Lesley E. Scott, Alexio Capovilla, Natela Rekhviasvili, Wendy Stevens. University of the Witwatersrand

A isothermal amplification termed Reverse Transcription Loop Dependant Amplification (RT-LDA) was developed with an affordable nucleic acid lateral flow detection (NALF) system, as one component of a potential POC HIV-1 RNA assay for subtypeC. RT-LDA makes use of a primer design that efficiently converts viral RNA into ssDNA amplicons, in 1 hour at 53ºC. Due to the single stranded nature of the product, the amplicon could be detected using NALF.

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