TaqMan® Endogenous Control Assays
A convenient collection of pre-designed TaqMan probe and primer sets enables you to normalize your gene expression data based on RNA sample amount. Endogenous control assays are available for the most commonly used human, mouse and rat genes, as well as 18S eukaryotic ribosomal RNA.
The goal of traditional real-time quantitative PCR gene expression experiments is to measure the concentration of specific RNA targets in each sample. Concentration requires knowledge of both the target amount and the size of the sample it was derived from. In this type of experiment, the most common method to measure sample size is to measure the RNA from a gene with consistent expression across all the samples in the experiment. Such genes have many names: normalizer genes, house-keeping genes, reference genes and control genes. TaqMan® Endogenous Control Assays are a collection of assays used in quantitative PCR gene eukaryotic expression experiments to measure sample size. Each assay consists of two primers and a TaqMan® probe that specifically amplifies and detects sequences derived from specific gene transcripts. TaqMan® Endogenous Control Assays are offered in multiple formats and scales for ease of use and economy.
Complete Solution for Quantitative Gene Expression
TaqMan® Endogenous Control Assays are part of the complete real-time PCR solution offered by Life Technologies™. The complete solution includes a wide variety of RNA extraction or cell lysis chemistries, multiple reverses transcription chemistries, the largest and highest quality collection of real-time PCR assays in the world, a variety of TaqMan® reagents, multiplexing support, instrumentation, plastics and software. TaqMan® Endogenous Control Assays fit neatly into the real-time PCR component, either in singleplex or multiplex.
Simple to Use
TaqMan® Endogenous Control Assays are simple to use. Each one works optimally under universal cycling and universal chemistry conditions. There is no optimization. The assays may be used with the One-Step, Two-Step or cell lysis RT PCR chemistries. Individual assays are simple to use: an assay master mix is created by adding a TaqMan® master mix, TaqMan® Endogenous Control Assay and water; each well receives assay master mix and sample. Real-time PCR arrays are even simpler to use: a sample - master mix mixture is dispensed to each well.
TaqMan® Endogenous Control Assays are offered in multiple formats and scales. Individual assays are formulated in a 20x liquid concentrate of two primers and a TaqMan® probe. Assay scales are large, medium and small. Large scale can be the most economical, because the same endogenous control assay can be used for many gene expression experiments. For example, each large scale assay is sufficient for 1000 PCR reactions at 50ul each or 2500 PCR reactions at 20ul each, making the cost per reaction very small. Another assay format is dry in real-time PCR arrays. Arrays are offered in multiple plate types: Fast 96-well plates, standard 96-well plates and TaqMan® Array Cards.
Choosing the Right Endogenous Control
Genes with fluctuating expression patterns should not be used as endogenous controls, because they can create large accuracy errors in the final results. Some endogenous controls may be accurate for certain sample types or tissues, but not others. If there is uncertainty in which endogenous control is most accurate for a specific sample set, endogenous control candidates may be tested in an easy, economical way using arrays. For example, TaqMan® Array Endogenous Control 96-Well Plates contain 32 candidate endogenous control assays, allowing up to 3 samples to be tested per plate. These plates may be purchased individually, maximizing flexibility to test as few or as many samples as needed. Once the candidate gene with the lowest variation has been identified from the array data, the individual TaqMan® Endogenous Control Assay for that gene may be purchased for the quantification experiment.
Multiplex vs. Singleplex PCR
Singleplex is a type of real-time PCR experiment in which only one gene is measured per well, while a multiplex experiment measures two or more genes per well. The most common type of multiplex is a duplex, where the target gene and control gene are measured in the same well. This type of multiplex is the easiest to validate and offers multiple benefits, including correcting for sample pipetting error, improved throughput and decreased sample usage. TaqMan® Endogenous Control Assays are offered in either singleplex format, which is a FAM™ labeled probe and universal primer concentration formulation, or multiplex format, which is a VIC® labeled probe and primer limited formulation. The VIC® labeled TaqMan® Endogenous Control Assay can be multiplexed with a FAM™ labeled TaqMan® Assay for the target gene. This dye combination works well on all Life Technologies™ real-time PCR instruments. Note that some TaqMan® Endogenous Control Assays contain a TAMRA™ quenched probe, which works on all Life Technologies™ real-time PCR instruments, except the StepOne™.
For Research Use Only. Not for use in diagnostic procedures.
This product is a Licensed Probe. Its use with an Authorized Core Kit and Authorized Thermal Cycler provides a license for the purchaser's own internal research under the 5’ nuclease patents and basic PCR patents of Roche Molecular Systems, Inc. and F. Hoffmann-La Roche Ltd. No real-time apparatus or system patent rights or any other patent rights owned by Applied Biosystems, and no rights for any other application, including any in vitro diagnostic application under patents owned by Roche Molecular Systems, Inc. and F. Hoffmann-La Roche Ltd claiming homogeneous or real-time amplification and detection methods, are conveyed expressly, by implication or by estoppel.
This product is optimized for use in the Polymerase Chain Reaction (PCR) process covered by patents outside the U.S. owned by F. Hoffmann-La Roche Ltd. No license under these patents to use the PCR process is conveyed expressly or by implication to the purchaser by the purchase of this product.
Life Technologies offers more than 1.3 million predesigned TaqMan® Gene Expression Assays covering 23 species, including some pathogens, the most comprehensive set of real-time PCR assays available.
You can order custom, all-in-one-tube TaqMan® reagent-based assays to perform gene expression studies on any gene or splice variant in any organism.
These instruments are economical, yet highly capable. Both may use Fast or standard ramp mode, use passive reference normalization and can multiplex. The StepOne™ features a fixed 48-well Fast block, while the StepOnePlus™ has a fixed 96-well Fast block.
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