12 Current white paper

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Molecular Probes® NanoOrange® Assay Performed on BMG LABTECH FLUOstar OPTIMA Microplate Reader

08-05-2008

The field of proteomics has expanded dramatically in recent years with research on a whole host of organisms. Techniques such as two dimensional difference gel electrophoresis (2D DIGE) require the accurate quantitation of protein, as up to three labelled samples can be loaded on a single gel for comparison. Assay methods for determining protein quantitation include absorbance at 280 nm(1), the Bradford assay(2), Lowry assay(3), BCA method(4) and more sensitive assays such as Fluoroprofile® (Sigma-Aldrich) and NanoOrange® (Molecular Probes®)5()....

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Real-Time Multiplex PCR of five Different DNA Targets Using the LightCycler® 480 System

19-09-2007

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

19-09-2007

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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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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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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Quantification of Zea mays mRNAs by Real-Time PCR Using the Universal ProbeLibrary

24-08-2007

The Universal ProbeLibrary (UPL) is a detection system for real-time PCR based on locked nucleic acid (LNA) probes. We tested the efficiency of the UPL for the quantification of maize transcripts on two detection platforms. The results indicate that the UPL is an attractive alternative to assays based on intercalating dyes or conventional probes....

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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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Genome Sequencer 20 System: Breakthrough in a New Applications Age of Sequencing

20-08-2007

The Genome Sequencer 20 System is an ultra-high-throughput automated DNA sequencing system capable of carrying out and monitoring sequencing reactions in a massively parallel fashion. Since the Genome Sequencer 20 System provides a complete solution for ultra-high-throughput DNA sequencing, an individual researcher can for the first time prepare samples and sequencing reactions, generate sequence reads, and assemble genome sequence data within days....

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Superiority of FuGENE® HD Transfection Reagent in Minimizing Non-Specific Cellular Interferon Responses

14-08-2007

Many viruses, including human cytomegalovirus (HCMV), encode for genes that can modulate or interfere with the ability of cells to mount an effective antiviral response to infection. Transfection of viral genes into cultures of mammalian cells is an important research tool to evaluate the function of viral gene products. We utilized this approach to identify viral genes involved in modulation of interferon (IFN)-mediated cell signaling pathways....

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