Real-time PCR Gene Expression Profiling Mikael Kubista, José Manuel Andrade, Björn Sjögreen and Amin ForootanCorrect interpretation of real-time PCR data requires appropriate experimental design, accurate data pre-processing and analysis of the data using proper statistical and multivariate methods. For this process we have developed the GenEx software. |  | |
Sensitive and Specific Detection of microRNAs Martin Kreutz, James Qin, Holger Engel, Po-Jen Shih, Peter Hahn, Martin Schlumpberger, Subrahmanyam Yerramilli and Eric LaderMicroRNAs are endogenous, 21–22 nt, noncoding RNAs that mediate post transcriptional gene regulation. miRNAs are involved in regulation of gene expression during development, differentiation, cell proliferation, and apoptosis. Misregulation of miRNA expression is associated with several cancers and other diseases. We have developed the miScript System for real-time PCR analysis of hundreds of miRNAs, sno RNAs, piRNAs, other small noncoding RNAs. |  | |
Real-time PCR in Diagnostics of Plant Viruses Mehle N., Boben J. and Ravnikar M.Sensitive and specific method real-time PCR was developed for diagnostics of certain plant viruses. Real-time PCR can be used for determination of different viruses as well as for differentiation between related strains of viruses. The method can also be used for detection of low concentrations of plant viruses in various samples such as environmental waters, growth substrates and seeds. |  | |
Real-Time Multiplex Rt-PCR on Circulating Tumor Cells Anieta M. Sieuwerts, Stefan Sleijfer, Jaco Kraan, Joan Bolt-de Vries, Jan-Willem Gratama, John WM Martens and John A. FoekensUsing the CTC kit (CellSearch™), cells that attached to anti-EpCAM Moab were immunomagnetically separated and used for analysis of a selected pilot set of 32 genes by real time RT-PCR. This study shows the feasibility of multiple gene expression analysis on RNA isolated from only one tumor cell. Most importantly, expression analysis of several tumor-specific genes in blood samples containing only 2 tumor cells is already possible. |  | |
Novel Fluidics Microbead Trap/Flow Cell Enhances Speed/Sensitivity of Bead-Based Bioassays Up to 5-Fold RM Ozanich, CJ Bruckner-Lea, JW Grate, MG Warner, BP Dockendorff, KC Antolick, HC Edberg, LH Johnson, AN EasterdayPacific Northwest National Laboratory (PNNL) has developed a micro/nano particle trap that allows surface-functionalized magnetic or non-magnetic particles to be trapped with subsequent perfusion of sample, reagents and wash solutions, yielding significant (up to 5-fold) improvements in assay speed and sensitivity, while significantly reducing sample matrix effects. |  | |
Utilizing High Speed Photography to Optimize Low Volume Dispensing Conditions Mary Cornett, Mitch Gordon and Anca RotheIn this study we use high-speed photography as a feedback mechanism for adjusting the Nanodrop instrument dispense settings to improve the positional dispense accuracy of low volume (nanoliter) drops. These same parameters can be investigated, with various fluid classes, to reduce deleterious effects on dispensing performance such as deflected streams, satellite formation, secondary pulses and drop deformation. |  | |
MicroRNA Profiling of Breast Cancer using miRCURY™ LNA Arrays Thomas Litman, Mikkel Nørholm, Christian Glue, Nina Stahlberg, Nana Jacobsen, Jens Eriksen, Inge M Svane, Henrik Flyger, Eva Balslev, Carsten Alsb and Søren MøllerMicroRNAs (miRNAs) comprise a recently identified class of small, non-protein-coding regulatory molecules that play important roles in many physiologic and pathologic processes, including differentiation, viral infection, and oncogenesis. In cancer, abnormal miRNA expression suggests that these molecules may serve as valuable diagnostic and prognostic molecular signatures. |  | |
Determination of Interferon-Pathway–Related Gene Induction during RNAi Jie Kang, Peter Hahn, Frank Narz, Cornelia Schmidt, Kirsten Haussuehl, Dong Liang, Subu Yerramilli, Eric Lader and Dirk Löffert,Experimental approaches using short interfering RNA (siRNA) molecules to specifically silence gene expression have become more widely used in recent years. As for all techniques in molecular and cellular biology, the importance of protocol optimization and the use of appropriate controls cannot be overestimated. |  | |
The BioRobot® Universal System - Multiple Applications on one Instrument Thomas Weierstall, Anja Schultz, Daniel Langendörfer, Martin Heller and Carola SchadeThe BioRobot Universal System enables standardized sample preparation and reaction setup using optimized, ready-to-run protocols. Fully tested protocols provide purification of RNA from blood, cells, and tissue and purification of genomic DNA from blood and buccal swabs, as well as RT-PCR and PCR setup. |  | |
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