19 Current white paper about the topic expression

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High sensitive detection of follistatin by Imperacer®: an initial study on the way to analytical gene-doping tests

08-05-2008

For the understanding of interactions in the muscle-growth directing myostatin-pathway and therefore also for the establishment of high sensitive detection strategies for gene-doping, detailed studies of protein concentration levels in biological matrices and appropriate highly sensitive analytic techniques for quantitative monitoring of these biomarkers are required. In this work, the development of a high sensitive assay for follistatin based on the Imperacer® technology is described. The quantification limit in human serum was found at 60 pg/ml in only 6 µl sample volume. With this initial assay we demonstrate the first application in a set of analytical tests for a high sensitive myostatin pathway related profiling....

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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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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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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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Efficient Transfection of Neuroblastoma Cell Lines Using FuGENE® HD Transfection Reagent

24-08-2007

Efficient transfection of cells is a prerequisite for genetic manipulation. However, many cell lines remain hard to transfect with established protocols, hampering consecutive analysis. Here, we compare three different methods (FuGENE® HD Transfection Reagent, transfection reagent L, and calcium phosphate) for the transfection of neuroblastoma cell lines. We show that FuGENE® HD Transfection Reagent is able to transfect all of these cell lines with high efficiency for further analysis....

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Superior Transfection of Human Embryonic Stem Cells with FuGENE® HD Transfection Reagent

21-08-2007

Gene manipulation techniques are potentially powerful tools in many applications for human embryonic stem (ES) cell research. Controlling expression levels of specific genes may improve the efficiency of differentiation induction. Cell types of interest may be collected using marker genes and antibiotic resistance genes. Tracking of ES cells derived after engraftment could be facilitated by tracer genes. It may also be possible to selectively destroy transplanted cells....

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Epitope Mapping Using the In Vitro Rapid Translation System RTS 100

21-08-2007

The Rapid Translation System (RTS) represents a cell-free system that allows efficient production of protein from linear DNA sequences. This article gives a brief outline of the use of the RTS technology in linear epitope mapping and shows its promise for reducing time-consuming cloning and cell-lysis steps....

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Analysis of the DNA Methylation Patterns at the BRCA1 CpG Island

20-08-2007

Germ-line alterations of the BRCA1 gene confer a lifetime risk of 40% for ovarian cancers and of 40%-80% for ­breast cancers. It is likely that BRCA1 acts as a tumor suppressor gene. BRCA1 involvement in breast cancers does not seem to be restricted to familial cancers. Despite the absence of somatic mutations in the breast tissues, a down regulation of BRCA1 expression is associated with malignancy in human sporadic breast cancers....

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Quantification of BRCA1 Expression Level Using Standard RT-PCR Reagents - a Sensitive Method for the Detection of Low Amounts of Transcripts

17-08-2007

Mutations in the breast cancer susceptibility gene, BRCA1, occur in the majority of families with multiple members affected with breast or ovarian cancer. Women who inherit a BRCA1 mutation have 40%-80% risk of developing breast cancer or ovarian cancer. However, up to 90% of breast cancers are sporadic, and mutations in BRCA1 seem to be a rare event in sporadic breast tumors....

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