Navigation Links
Amplification Efficiency of TaqMan Gene Expression Assays

u and Saint, 20022. The calculated fold change from each method was plotted versus the expected fold change, and the correlation between calculated and expected results was determined.

Results

Determination of Amplification Efficiencies of TaqManGene Expression Assays

Initial testing of 30 assays using a 1-log dilution range (with three concentration points at 10, 5, and 1 ng/μL) and a 3-log dilution range (with four concentration points at 10, 1, 0.1, and 0.01 ng/μL) indicated there were statistical problems with these narrow dilution ranges when measuring amplification efficiencies. Narrow dilution ranges are often used because of the limited amount of cDNA available to do the experiment, especially when quantitating low-copy number transcripts within a cDNA sample.

The use of a broad dynamic range and more data points significantly altered the distribution of amplification efficiency values. The use of a broader log dilution range, however, requires the generation of PCR product or other high concentration template such as a cDNA clone. Therefore, the best protocol for determining amplification efficiency is more difficult to implement.

The initial data indicated that the accuracy of efficiency measurements is extremely sensitive to both the number of data points and the dilution range used in a single experiment. Replicate experiments indicated that the precision of an amplification efficiency measurement increases when the experiment is performed over a broader range of template concentrations, or dilutions.

To further define the variability associated with measuring amplification efficiencies, we tested one assay targeting the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) transcript using either a 2-log (cDNA template) or 5-log (PCR product) dilution series with 5 and 6 concentration points, respectively. M
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
2. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
3. Greater Amplification Specificity with New Hot Start PCR Enzyme
4. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
5. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
6. Improve Amplification Specificity with Hot Start PCR Enzyme
7. Amplification of the amoA Gene of Nitrosococcus oceani Bacteria Using Eppendorf MasterTaq
8. Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
9. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
10. Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler
11. Amplification of Epstein-Barr Virus Exon C with Difficult Primers Using Eppendorf Mastercycler gradient
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Amplification Efficiency TaqMan Gene Expression Assays

(Date:8/22/2014)... first direct observations of how facets form and ... more sophisticated and effective nanocrystal design and reveal ... crystal growth breaks down at the nanoscale. , ... Lawrence Berkeley National Laboratory (Berkeley Lab) used highly ... fast-detection camera to capture the physical mechanisms ...
(Date:8/22/2014)... Aug. 22, 2014 Research and Markets ... and China Gel Permeation Chromatography (GPC) Industry Report 2014" ... Global and China Gel Permeation Chromatography (GPC) Industry Report ... state of the global gel permeation chromatography industry with ... provides a basic overview of the industry including definitions, ...
(Date:8/22/2014)... August 22, 2014 The identification ... organisms can be accelerated by exome sequencing. Built ... technologies, the custom SeqCap EZ Developer system has ... Mr. Watson will additionally present how he achieved ... available human and mouse exome kits. With over ...
(Date:8/21/2014)... Finnish-Swedish research group at the Institute for Molecular ... institutet, Stockholm, has developed a novel "man and ... This innovative diagnostic aid was described in ... The method is based on computer vision algorithms ... combined with visualization of only the diagnostically most ...
Breaking Biology Technology:Shaping the future of nanocrystals 2Shaping the future of nanocrystals 3Shaping the future of nanocrystals 4Global and China Gel Permeation Chromatography (GPC) Industry Report 2014 2Efficient Identification of Protein-Coding Variants in a Model Organism Through Exome Sequencing, New Webinar Hosted by Xtalks 2A novel 'man and machine' decision support system makes malaria diagnostics more effective 2
... in Latin,America, WASHINGTON, June 12 The ... San Francisco (UCSF) announced,today that 10 promising biomedical ... in the Biomedical Sciences. Funded by Pew through ... offers talented,young Latin American scientists $60,000 for a ...
... today the,results of tests utilizing its Ygiene(TM) formulation ... is designed to be used by the,military and ... "The Ygiene(TM) formulation killed all anthrax spores exposed ... said Dr. Philip Tierno, the,Principal Investigator on the ...
... Company Begins Process to Restore Shareholder Value Following NEBIDO(R) ... ... Indevus,Pharmaceuticals, Inc. (Nasdaq: IDEV ) today announced that Glenn L. Cooper,MD, ... the Company through the challenges,it faces as a result of the recent ...
Cached Biology Technology:Ten Latin American Scientists Named 2008 Pew Fellows in the Biomedical Sciences 2BioNeutral Announces a BREAKTHROUGH in the Fight Against Bioterrorism 2Indevus Announces Management Changes 2Indevus Announces Management Changes 3Indevus Announces Management Changes 4
(Date:8/22/2014)... of captive black rhino reproduction in Europe highlights ... breeding programmes. , Researchers from Chester Zoo, The ... carried out a six-year study which encompassed 90% ... Katie Edwards led the research as part of ... says: "Although some black rhinoceros breed well in ...
(Date:8/22/2014)... and the bees. But that old expression leaves ... fertilization process for flowering plants is particularly complex ... female reproductive cells. New research from an international ... including Carnegie,s Wolf Frommer, David Ehrhardt, and Guido ... that guides flowering plant fertilization. It is published ...
(Date:8/21/2014)... -- Nxt-ID, Inc. (OTCQB: NXTD), a biometric authentication company ... that its shares of common stock and the warrants ... proposed underwritten public offering of common stock and warrants ... Market, subject to closing of its proposed underwritten public ... trade under the symbol "NXTD" and "NXTDW," respectively. ...
Breaking Biology News(10 mins):Hormone analysis helps identify horny rhinos 2Calcium and reproduction go together 2Nxt-ID Approved for NASDAQ Capital Market Listing 2Nxt-ID Approved for NASDAQ Capital Market Listing 3
... forests of Thailand have given conservationists confirmation that anti-poaching ... the Wildlife Conservation Society. The footagetaken by ... Complex over the last yearhas revealed a cornucopia of ... gaurs, sun bears, and many other species. The clips ...
... Researchers with the U.S. Department of Energy (DOE),s ... of time-lapse live imaging and mathematical modeling of a ... to suggest that for low dose levels of ionizing ... dose. This contradicts the standard model for predicting biological ...
... human skulls have shown that changes to the skull ... events may have actually precipitated each other. Researchers ... skulls from the Austrian town of Hallstatt and found ... variation in one part of the skull is linked ...
Cached Biology News:Candid video clips from Thailand show anti-poaching efforts saving wildlife 2New take on impacts of low dose radiation 2New take on impacts of low dose radiation 3New take on impacts of low dose radiation 4Human skull study causes evolutionary headache 2
... its Best: DASGIPs Process Control integrates ... & comprehensive documentation for cultivation processes. ... as pH, dissolved oxygen, temperature and ... online. Profiles, e.g. constant or scheduled ...
... Microarrays designed for immunological studies, vaccine ... the flexible and powerful Paraflo microfluidic ... available as part of our comprehensive ... Custom Probe Content Probe content ...
... are custom designed to meet your needs. ... 8,455 features per array (2 arrays/1" x ... array (1 array/slide). Features ... - This extended length delivers higher sensitivity ...
... Agilent Two-Color RNA Spike-In Kit contains in ... only to complementary control probes on Agilents ... spike-ins serve as a positive microarray control ... NOTE: This kit is a required component ...
Biology Products: