11 Current white paper

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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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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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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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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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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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siRNA Silencing of Lamin A and Quantification of the Knockdown Effect via qPCR

08-08-2007

Lamins are structural proteins which form a thin fibrous layer called the nuclear lamina. Currently, three types of lamins are known in mammalian cells: lamin A, B, and C. Lamins may be involved in DNA replication, chromatin organization, differentiation, nuclear structural support, nuclear envelope reassembly, and several other processes. Mutations of lamin A can cause muscular dystrophy, dilated cardiomyopathy, familiar partial lipodystrophy [3], and other disorders. For the detailed analysis of the different functions of this more than 70 kDa large protein, the expression should be blocked....

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DNase I Recombinant, RNase-free

High-Quality Tool for Demanding Applications

06-08-2007

Preparation of RNA, free from DNA, is a critical step before performing RT-PCR assays. Total RNA isolated from several sources using routine methodologies is frequently contaminated with DNA that can give rise to amplification products, which mimic the amplicons expected from RNA targets. DNase I recombinant, RNase-free safely and efficiently eliminates genomic DNA contaminations from RNA samples....

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cDNA Synthesis with the Transcriptor First Strand cDNA Synthesis Kit from Formalin-Fixed Paraffin-Embedded Tissue (FFPET) Samples

11-06-2007

Partial degradation of RNA is a common problem when working with FFPET samples. Semi-­quantitative RT-PCR assays amplifying the 5´- and 3´-end sequences of the housekeeping gene b-actin present a solution: calculating the ratio of the 3´- to 5´-amplicon allows ­assessment of RNA integrity. An important step for reproducible quantification is the reverse ­transcription reaction. The ­applicability of the new Transcriptor First Strand cDNA Synthesis Kit to partially degraded FFPET RNA was evaluated. The new kit is compared with the widely used First Strand cDNA Synthesis Kit for RT-PCR (AMV). Under modified reaction ­conditions (temperature, time) the kit reversely transcribes the FFPET samples with high ­efficiency....

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