Navigation Links
Amplification Efficiency of TaqMan Gene Expression Assays

p> Universal PCR Master Mix without AmpErase UNG on an ABI PRISM 7900HT Sequence Detection System. The CT values obtained over this 6-log dilution range were plotted against cDNA concentration (Figure 1). Dilutions with a CT higher than 35 were not included in the results because CT values greater than 35 approach the sensitivity limits of the real-time PCR detection system. Amplification efficiency was calculated from the slope of this graph using the equation:

Equation 3

For example, if the slope of the line is -3.33, then the amplification efficiency is:

Use of the Comparative CT Method for Determination of Fold Change in Gene Expression

Stratagene Universal Human Reference RNA (100 ng/μL) was reverse transcribed as previously described to obtain cDNA. The cDNA was diluted in a 100-fold dilution range to obtain a dilution series with known amounts of cDNA. A constant amount of cDNA reverse transcribed from a nonhuman transcript (not detected in the Stratagene RNA pool) was used as a control to measure fold change. This transcript was made by T7 transcription of a 227 bp fragment from the Hepatitis C virus 5' UTR inserted into a TOPO-TA plasmid. Each of the Universal Human Reference cDNA dilutions was subjected to real-time PCR amplification as described above, using TaqMan Universal PCR Master Mix without AmpErase UNG on an ABI PRISM 7900HT Sequence Detection System. The relative quantities of diluted target cDNA vs. Hepatitis C cDNA were determined by either the nnCT method (Applied Biosystems User Bulletin #23) or by inserting the pre-determined efficiency values for target and endogenous control into the equation:

Equation 4

For derivation of the above equation, see Li
'"/>

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:9/30/2014)... Over the last decade, the laboratory filtration ... budgets and increased acceptance of new filtration technologies in ... industries are also catalyzing the growth of laboratory filtration ... to grow at a strong CAGR during the forecast ... $2,532.6 million by 2019. The market is mainly driven ...
(Date:9/30/2014)... 30, 2014 Follow us on ... each type of cholesterol carrier or lipoprotein in the ... excessive cholesterol levels in the body, especially along the ... heart attack and other cardiovascular disorders. Cardiovascular Diseases (CVD) ... for cholesterol levels on a regular basis, to assess ...
(Date:9/30/2014)... 2014 CORD:USE Cord Blood Bank – ... has entered into an Equity and Exclusive Services Agreement ... series of patented technologies for the isolation of human ... Tianhe is performing Phase I/II clinical trials in ... using Tianhe,s Stem Cell Educator Therapy showing that ...
(Date:9/30/2014)... Genedata, a leading provider ... and related life science research, today announced ... Selector as its host-pathogen data analysis platform ... a European consortium dedicated to improving the ... anti-fungal drug therapies. Genedata Selector supports this ...
Breaking Biology Technology:Global Laboratory Filtration Market To See 7.0% CAGR to 2019 Says a Research Report Available at ReportsnReports.com 2Global Laboratory Filtration Market To See 7.0% CAGR to 2019 Says a Research Report Available at ReportsnReports.com 3Global Laboratory Filtration Market To See 7.0% CAGR to 2019 Says a Research Report Available at ReportsnReports.com 4Global Laboratory Filtration Market To See 7.0% CAGR to 2019 Says a Research Report Available at ReportsnReports.com 5Surging Diabetic, Obese and Hypertensive Population Drives the Global Cholesterol Testing Market, According to New Report by Global Industry Analysts, Inc. 2Surging Diabetic, Obese and Hypertensive Population Drives the Global Cholesterol Testing Market, According to New Report by Global Industry Analysts, Inc. 3Surging Diabetic, Obese and Hypertensive Population Drives the Global Cholesterol Testing Market, According to New Report by Global Industry Analysts, Inc. 4CORD:USE is Pleased to Announce it has Partnered with Tianhe Stem Cell Biotechnologies, Developer of Cord Blood Stem Cell Therapy Showing Promising Results in Clinical Trials for the Treatment of Diabetes and Autoimmune Diseases 2CORD:USE is Pleased to Announce it has Partnered with Tianhe Stem Cell Biotechnologies, Developer of Cord Blood Stem Cell Therapy Showing Promising Results in Clinical Trials for the Treatment of Diabetes and Autoimmune Diseases 3CORD:USE is Pleased to Announce it has Partnered with Tianhe Stem Cell Biotechnologies, Developer of Cord Blood Stem Cell Therapy Showing Promising Results in Clinical Trials for the Treatment of Diabetes and Autoimmune Diseases 4FUNGITECT Consortium Adopts Genedata Selector as Data Analysis Platform for Anti-fungal R 2FUNGITECT Consortium Adopts Genedata Selector as Data Analysis Platform for Anti-fungal R 3
... Wisconsin-Madison , justified or otherwise, is that its academic ... has world-class scientists and well-developed mechanisms for turning their ... that commercialization takes place within a half-hour drive from ... the figures for per capita wages, unemployment and job ...
... subscribe to the theory that consumer need is what drives ... some proof. , ,In this case, the need is ... something to be desired, and the chief executive of a ... and broadband problems before. , ,Albert Chiozzi, who plans ...
... half of the nation's top 25 banks have joined an ... , a business unit of Metavante Corp ., the ... ,Twelve of the nation's largest banks now will be able ... physically convey them, a capability that was encouraged by the ...
Cached Biology Technology:Entrepreneurship grant will build strong foundation 2Entrepreneurship grant will build strong foundation 3Entrepreneurship grant will build strong foundation 4Company to operate in wireless Internet space 2
(Date:9/30/2014)... Prenatal exposure to tetrachloroethylene (PCE) in drinking water ... abruption, according to a new study led by ... , The study, published in the journal ... 1,019 unexposed pregnancies among 1,766 women in Cape ... late 1960s to the early 1980s by the ...
(Date:9/30/2014)... -- Pollution in urban and farm runoff in Hawaii ... study finds. , The study, published Tuesday in ... nitrogen in the runoff ends up in algae that ... the animals, eyes, flippers and internal organs. , Scientists ... National Oceanic and Atmospheric Administration (NOAA) conducted the study ...
(Date:9/30/2014)... trouble getting pregnanteven with IVF? Here,s some hope: A ... The FASEB Journal , explains how ... known as PAWP, which induced embryo development in human ... embryo development by the sperm cell during fertilization. , ... a major paradigm shift in our understanding of human ...
Breaking Biology News(10 mins):Contaminated water linked to pregnancy complications, BU study finds 2Pollution linked to lethal sea turtle tumors 2Pollution linked to lethal sea turtle tumors 3Synthetic sperm protein raises the chance for successful in vitro fertilization 2
... may have to exercise even harder to get the benefits ... in the February issue of the journal Cell Metabolism, published ... a kind of metabolic boost in response to exercise is ... AMP-activated protein kinase (AMPK) slows down in the skeletal muscle ...
... Having a twin sister could put male saiga antelopes at ... shows that male twins with a sister are born lighter ... optimal size for males. The research also shows that saigas ... other similar species investing more in their offspring during ...
... how much they had already eaten -- in this case, ... who had no such environmental cues, finds a new Cornell ... eating at a table where leftover bones accumulated compared with ... than for women. , "The results suggest that people ...
Cached Biology News:Power-boosting signal in muscle declines with age 2The penalty of having a sister -- why sibling sex matters for male saiga antelopes 2The penalty of having a sister -- why sibling sex matters for male saiga antelopes 3