22 Current white paper about the topic genes

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Efficient Transfection of Primary Human Skeletal Myoblasts Using FuGENE® HD Transfection Reagent

26-09-2007

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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Mutation Scanning of the Cytidine Deaminase Gene by High-Resolution Melting Curve Analysis Using the LightCycler® 480 System

12-09-2007

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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Genotypes in Smokers: Correlations with Smoking Behavior

12-09-2007

Smoking behavior is influenced by both genetic and environmental factors. Moreover, smoking initiation and smoking persistence have a heritability of at least 50%. A large body of work has been dedicated to associating multiple genetic markers of neurobiological pathways previously linked to addiction and smoking such as the central dopaminergic, serotoninergic, and nicotinergic pathways with different aspects of smoking behavior. In order to investigate the feasability of future genetic analyses, we genotyped 14 SNPs in 288 samples of addicted smokers using the LightCycler® 480 System with HybProbe assays....

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Metagenomics Analysis Using the Genome Sequencer FLX System

05-09-2007

The Genome Sequencer FLX System from 454 Life Sciences and Roche Applied Science is a versatile sequencing platform suitable for a wide range of applications, including de novo sequencing and assembly of genomic DNA, transcriptome sequencing, small RNA analysis, and amplicon sequencing. The Genome Sequencer FLX is built upon 454 Sequencing technology that enables long read lengths and very high single-read accuracy. One application in which the technology is accelerating the field's understanding is metagenomics....

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Real-Time PCR Quantification of Plant miRNAs Using Universal ProbeLibrary Technology

29-08-2007

MicroRNAs (miRNAs) are naturally occurring, highly conserved families of transcripts (18-25 nt in length) that are processed from larger hairpin precursors. Both plant and animal genomes encode miRNAs, and to date, there are over 4000 mature miRNA transcripts annotated. Although miRNAs represent a relatively abundant class of transcripts, their expression levels vary greatly among species and tissues. Less abundant miRNAs routinely fail detection using technologies such as cloning, northern hybridization and microarray analysis. Here, we present a real-time quantification method for accurate, sensitive, and economical detection of plant miRNAs based upon the stem-loop RT primer approach using an miRNA-specific primer coupled with a stem-loop specific Universal ProbeLibrary (UPL) probe. This approach represents a highly specific, sensitive, and easy-to-apply system to detect miRNAs....

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The Universal ProbeLibrary - a Versatile Tool for Quantitative Expression Analysis in the Zebrafish

29-08-2007

The teleost zebrafish is a widely used model organism for the study of developmental and disease-related processes in vertebrates, including humans. Understanding these processes at a molecular and cellular level frequently requires the estimation of gene expression levels. Quantitative real-time PCR (qPCR) is an important tool towards this goal but has, until now, been relatively neglected in zebrafish. Here, we describe the comparison of the Universal ProbeLibrary (UPL)-based qPCR to a conventional SYBR Green I-based qPCR assay using embryonic zebrafish cDNA. We found that the Universal ProbeLibrary-based assay shows equal or better performance compared with the SYBR Green I assay....

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Real-Time PCR Quality Control for Gene Expression

Profiling Using the LightCycler® 480 System

28-08-2007

Quantitative real-time PCR (qPCR) has become the de facto standard for nucleic acid quantification. This achievement is due in large part to its sensitivity, specificity, accuracy, large dynamic range of linear quantification, and its speed. The qPCR technology has matured to a ready-to-use commonly available method in most molecular biology laboratories. Nevertheless, the reliability of the final quantification result depends heavily on all elements in the workflow, such as the quality of the input template (integrity and absence of inhibitors), the PCR assay (specificity, efficiency, limit of detection), and normalization strategy (validated reference genes)....

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The New Genome Sequencer FLX System

More flexibility, more breakthrough applications - Roche continues to unlock the sequencing secrets that will change the world

27-08-2007

In 2005, the Genome Sequencer 20 System was introduced into the global life science market. Based on the revolutionary 454 sequencing technology, this system is able to generate hundreds of thousands of sequence reads in a few hours at a fraction of costs compared with the traditional Sanger technology....

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Enabling Cancer Research with FuGENE® HD Transfection Reagent

27-08-2007

A major problem in some areas of cancer research is the difficulty of transfecting a significant number of the cells of interest, while minimizing the cytotoxic or other off-target effects of the transfection event itself. It is well-known that the use of some transfection reagents results in high levels of expression of the gene of interest in surviving cells, but also in high levels of cell death or other cellular alterations. FuGENE® HD Transfection Reagent overcomes these problems, results in high ­levels of transfection in many cancer cell lines, and thus enables researchers to conduct experiments that have previously not been possible....

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Ultrafast De Novo Sequencing of Corynebacterium urealyticum Using the Genome Sequencer 20 System

22-08-2007

A microbial genome sequence provides a wealth of data and specific information that cannot be obtained by other experimental approaches. A genome sequence can be regarded as the starting point for detailed bioinformatics analysis, metabolic reconstruction and systematic functional examination of all identified genes. In particular, genome sequencing has revealed important information on genes and deduced proteins of human pathogens, including putative virulence factors and potential new drug targets....

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