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Article 1 - 5 of 39 of the company Roche Diagnostics GmbH |
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Efficient Transfection of Primary Human Skeletal Myoblasts Using FuGENE® HD Transfection Reagent
Transfection of cells is one of the main techniques used to influence gene expression. Most primary cells and human skeletal myoblasts (SkMC) in particular are very difficult to transfect, whereas for cell lines such as C2C12, many suitable transfection reagents and protocols are available. Few publications report successful transfection of human primary myoblasts using non-viral systems [1,2,3]. These methods include cationic lipids such as phosphonolipids, electroporation, and a combination of liposome and adenoviral associated proteins. But transfection efficiency is low and often is a compromise between toxicity of the reagents and transfection efficiency....
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Enzymatic Protein Digest for Mass-spectrometric Analysis
The main focus in proteomic studies is on the identification of proteins in given biological samples. Proteins isolated and separated from a given sample (e.g., whole cell lysates, blood or tissue, protein complexes) by immunoprecipitation or affinity chromatography or two-dimensional electrophoresis must be proteolytically cleaved in the course of sample preparation for identification and characterization. A reproducible cleavage pattern of digested proteins is a prerequisite for the unambiguous identification of these proteins by mass spectrometry (MS)....
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Real-Time Multiplex PCR of five Different DNA Targets Using the LightCycler® 480 System
One of the most interesting aspects of real-time PCR based on detection of fluorophoric-labeled oligonucleotides, such as Hydrolysis probes, and Molecular Beacons, is the possibility of being able to detect conveniently multiple targets in the same PCR reaction (multiplex PCR). Ideally, a real-time multiplex PCR should be able to detect, differentiate, and provide a quantitative result for many different targets without a single target influencing the detection of one of the others (cross-talk) and without loss of sensitivity. It is evident that due to the limited number of fluorophoric labels available and the significant overlap in their emission spectra, quantification of multiplex reaction products is difficult and often not possible for more than two targets....
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High Pure PCR Cleanup Micro Kit - Novel Flexibility for Nucleic Acid Purification
Manual nucleic acid isolation with High Pure products is long approved by the scientific community. The product portfolio includes kits for isolation of genomic RNA and DNA, plasmid DNA as well as purification of reaction products. Key goal for all products is to offer optimal solutions suited for many different sample materials in order to reduce the number of different nucleic acid isolation kits necessary in modern molecular biology labs....
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Mutation Scanning of the Cytidine Deaminase Gene by High-Resolution Melting Curve Analysis Using the LightCycler® 480 System
Alexandre Evrard1,2, Caroline Raynal1,2, Jean-Christophe Boyer2, Lionel Le Gallic2, and Serge Lumbroso1,2 1Institut de Génomique Fonctionnelle, Département d'Oncologie Moléculaire et Cellulaire, Montpellier, France, 2Laboratoire de Biochimie, Hôpital Carémeau, CHU Nîmes, France *Corresponding author...
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Roche Diagnostics GmbH Mannheim, Germany
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