Reference cDNA for real-time qPCR
Measurements of mRNA expression levels—whether by Northern analysis, ribonuclease protection, or real-time quantitative PCR—are usually standardized by comparing the data to that obtained for an internal or endogenous reference gene. Housekeeping genes such as beta-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are most often used because their expression levels are expected to remain constant under different treatment conditions. Unfortunately, this assumption is not always valid, and results based on housekeeping genes alone can be biased (Suzuki, Higgins, and Crawford 2000). A better method is to normalize your data using our qPCR Human Reference cDNA, the only cDNA control derived entirely from human tissues.
Measurements of mRNA expression levels—whether by Northern analysis, ribonuclease protection, or real-time quantitative PCR—are usually standardized by comparing the data to that obtained for an internal or endogenous reference gene. Housekeeping genes such as beta-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are most often used because their expression levels are expected to remain constant under different treatment conditions. Unfortunately, this assumption is not always valid, and results based on housekeeping genes alone can be biased (Suzuki, Higgins, and Crawford 2000). A better method is to normalize your data using our qPCR Human Reference cDNA, the only cDNA control derived entirely from human tissues.
qPCR Human Reference cDNA is the ideal control for comparing data from different quantitative PCR (qPCR) experiments. Because it is prepared from a total RNA pool collected from several different tissues, our qPCR Human Reference cDNA provides broad gene coverage, as shown by microarray analysis of the RNA starting material. RNA, and therefore cDNA, prepared from whole tissues provides better gene representation with less variation than RNA made from cell lines (data not shown). Moreover, PCR analysis shows that our total RNA is virtually free of genomic DNA. This allows for a more accurate measurement of transcript copy number. Both high- and low-abundance genes are well represented, allowing preparation of a wide range of serially diluted standards for each qPCR assay. Lot-to-lot variation of reference cDNA is minimal because the RNA source is prepared on an industrial scale.
Overview
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Broad gene coverage
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Virtually free of genomic DNA
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Made from human tissues, not cultured cell lines
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A high-performance standard for quantitative PCR
More Information
Applications
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Real-time PCR/qPCR
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Gene expression analysis
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One-step RT-PCR
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Northern analysis
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Microarray analysis
Reference
Suzuki, T., Higgins, P. J. & Crawford, D. R. Control selection for RNA quantitation. Biotechniques 29, 332–7 (2000).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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