Bio-Rad Paper Describes the Effect of RNA Degradation on Data Quality in Quantitative PCR and Microarray Experiments

22 Nov 2007

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Bio-Rad Laboratories, Inc., a multinational manufacturer and distributor of life science research and clinical diagnostic products, today announced the availability of a technical paper (bulletin 5452A) that describes the effect of RNA degradation on data quality in quantitative PCR and microarray experiments. The paper describes using automated electrophoresis to determine RNA quality prior to gene expression analysis.

Quantitative expression levels, when determined by qPCR, decrease as RNA becomes increasingly degraded. This can lead to erroneous conclusions regarding the levels of gene expression when comparing samples that are degraded to different extents. Genes with high levels of expression can still be characterized but degradation compromises the ability to detect differences in genes that are expressed at low levels.

Bio-Rad’s Experion™ automated electrophoresis system is an effective method to determine the quality and quantity of RNA in gene expression analysis experiments. The Experion system accurately quantitates and evaluates the integrity of samples using as little as 25 ng total RNA, which is several hundred times less material than that required for gel electrophoresis. Once RNA quantity and quality have been established, a researcher can determine whether or not to move forward with additional experiments, knowing to what degree the results may be compromised.

The six-page paper provides a detailed introduction, methods description, results, and a discussion, supported by numerous graphs, tables, figures, and electropherograms.

Availability
Bulletin 5452A is available either from a local Bio-Rad sales office or it may be downloaded via the article webpage.

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ElectrophoresisElectrophoresis equipment is used to separate mixtures of protein, DNA or RNA, based on their electric charge, size and other physical characteristics, by passing them through a medium such as a polyacrylamide gel, an agarose gel or a capillary tube. Electrophoresis equipment includes horizontal and vertical gel electrophoresis chambers, isoelectric focusing systems, 2D electrophoresis and capillary electrophoresis instruments. Precast gels with a gradient can be used or gels can be hand cast. Samples are run alongside ladders or markers to identify the approximate size of a molecule. Separated proteins and nucleic acids in the gel can be stained or probed with fluorescent markers and then imaged and detected using gel documentation instruments, transilluminators, densitometers and scanners. Find the best electrophoresis equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Genome AnalysisGenomics, the study of genomes, includes functional genomics, evolutionary genomics and comparative genomics. There are many genomic technologies such as DNA sequencing of whole genomes, computational biology and bioinformatics. DNA and nucleic acids must be isolated and concentrated from cells for analysis with kits, automated analyzers and software. Other useful technologies for studying genomics include PCR, microarrays and electrophoresis.PCR and Thermal CyclingPolymerase chain reaction (PCR) kits and thermal cyclers are used for the in vitro amplification of DNA permitting subsequent analysis and experimental procedures. Explore a range of high-quality polymerase, primers and nucleotides or simplify your workflow with a PCR mastermix. Find reverse transcription PCR (RT-PCR) and cDNA synthesis kits for RNA products and libraries. Quantitatively measure the amplification of DNA with real-time PCR (qPCR) and droplet digital PCR (ddPCR) kits and systems, and discover automated PCR setup solutions to increase throughput. Alternative DNA amplification methods also include recombinase polymerase amplification (RPA) kits. Find the best PCR kits and thermal cyclers and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Microarray AnalysisMicroarrays, also known as biochips, are used for the detection and analysis of multiple genes, proteins, antibodies, or biomarkers on a single microchip. This can reveal information on protein or gene expression, single nucleotide polymorphism (SNP), copy number variation (CNV), epigenetics and patient health in clinical diagnostic tests. Discover a range of microarray scanners and prefabricated antibody, protein, RNA and DNA microarrays for your analysis or consider creating your own custom microarrays with a microarray printer. Find the best microarray products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.
Bio-Rad Paper Describes the Effect of RNA Degradation on Data Quality in Quantitative PCR and Microarray Experiments