Navigation Links
Real-Time PCR: General Considerations, Rev A

s clearly depicted in Figure 3. The threshold cycle was shifted by 4.7 cycles at 104 copies and by 3.6 cycles at 107 copies when the MgCl2 concentration was increased from 1.5 mM to 3.0 mM. This significant shift in CT was not observed in the 2.25 mM or 4 mM MgCl2 samples.

PCR efficiency can be affected by numerous factors. In this technical note we have evaluated 2 of these factors secondary structure interference and free magnesium ion concentration. We have clearly demonstrated that optimization of PCR design and reaction conditions on this template has strong effects on the quality of real-time PCR assays.

Once the primer locations and the MgCl2 concentrations have been evaluated, other reaction components, such as probe concentration and protocol temperatures, may need to be evaluated. For example, a 2-step or 3-step PCR thermal protocol may yield optimal amplification detection, depending on the detection strategy. Although this initial optimization may seem cumbersome, it will result in reaction conditions that are robust and reproducible.

* SYBR is a trademark of Molecular Probes, Inc. Practice of the patented polymerase chain reaction (PCR) process requires a license. The iCycler iQ system includes a licensed thermal cycler and may be used with PCR licenses available from PE Corporation. Its use with authorized reagents also provides a limited PCR license in accordance with the label rights accompanying such reagents. Some applications may require licenses from other parties.


back to top
'"/>

Source:


Page: All 1 2 3 4 5 6 7

Related biology technology :

1. Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System
2. Monitoring Microbial Populations Using Real-Time qPCR on the MJ Research Opticon 2 System
3. Rapid SNP Genotyping of the Pharmacogenomically Important Allele CYP2D6*4 Using Real-Time qPCR on the DNA Engine Opticon 2 System
4. Real-Time Multiplex PCR from Genomic DNA Using the iCycler iQ Detection System
5. Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A
6. Real-Time Quantification of Genomic DNA Using DyNAzyme II DNA Polymerase and SYBR Green I Dye
7. Quantitation of Lymphangiogenesis Using the iCycler iQ Real-Time PCR Detection System and Scorpions Detection System, Rev A
8. Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A
9. Real-Time Immuno-PCR on the iCycler iQ System, Rev A
10. Real-Time PCR/Melt-Curve Analysis: SNP Detection With FRET, Rev A
11. General Notes on Primer Design in PCR*
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Real Time PCR General Considerations Rev

(Date:9/15/2014)... Bioptigen announced today the availability of ... optical device metrology and pre-clinical ophthalmic research markets. ... Contact Lens Metrology is designed for non-destructive, non-contact, full ... IOL structures, and donor tissues. The metrology ... in a hydration bath (saline), automated segmentation of anterior ...
(Date:9/15/2014)... AUSTIN, Texas , Sept. 15, 2014 The ... changes in climate, says a new article published by the ... Ahmed , PhD, on climate change and its effects on ... part of an extensive study conducted by Dr. Ahmed in ... China and has implications for the future ...
(Date:9/15/2014)... Maryland , 15 de septiembre de 2014 ... se llevará a cabo del 12 al 18 de octubre. ... (PPTA) y de sus compañías miembro s, la ... la concienciación mundial en torno a la colección de fuentes ... donantes de plasma al salvar y mejorar las vidas ...
(Date:9/14/2014)... variety of ways, including by modifying the protein structure. ... form and perform no or even the "wrong" function: ... diseases such as Alzheimer,s, Parkinson,s or cystic fibrosis. Some ... other molecules of the same type and congregate into ... damage cells and tissues and make people ill. , ...
Breaking Biology Technology:Bioptigen Introduces Deep Imaging OCT System for Contact Lens Metrology 2Tea Taste and Quality Affected by Climate Change 2Tea Taste and Quality Affected by Climate Change 3Tea Taste and Quality Affected by Climate Change 4La International Plasma Awareness Week será una celebración para los donantes 2La International Plasma Awareness Week será una celebración para los donantes 3Measuring modified protein structures 2Measuring modified protein structures 3
... material that could become essential to efficient, environmentally ... advantage of graphene,s outstanding strength, light weight and ... wells. The Rice University lab of chemist ... Texas-based supplier of drilling fluids and subsidiary of ...
...  Kensey Nash Corporation (Nasdaq: KNSY ) today ... (Nasdaq: SPNC ) that they reached a ... product lines purchased from Kensey Nash in May 2008. ... payable to Kensey Nash.  As previously disclosed, the Company ...
... Sangui enters commercialization phase / Initial order of ... Tamaulipas has issued an initial order of Hemo2Spray, the Sangui ... product Scabremove both registered by the Mexican authorities in September ... the wound spray and Sangui management is positive that an ...
Cached Biology Technology:Functionalized graphene oxide plays part in next-generation oil-well drilling fluids 2Functionalized graphene oxide plays part in next-generation oil-well drilling fluids 3Mexican State of Tamaulipas Starts Training Doctors in Sangui Wound Management 2
(Date:9/16/2014)... years after the Hesperides vessel returned to Spain culminating ... have an increasingly clear picture of how the global ... the input of pollutants from the atmosphere is not ... most remote areas of the planet, and it is ... are presented this week in CSIC Residence for Researchers ...
(Date:9/16/2014)... . CRG researchers ... Journal of Cell Biology describes how Topo 2 ... , At this very moment thousands of our body,s cells ... the body repairs damaged tissues and regenerates others like skin ... "mitosis", during which the cell duplicates its genetic material and ...
(Date:9/16/2014)... Sept. 16, 2014  Cross Match Technologies, a ... announced today the launch of its Verifier ® ... identity of an individual using their secure credentials ... rapidly reads credential documents with embedded biometric data ... the biometric to a live scan of their ...
Breaking Biology News(10 mins):The Malaspina Expedition confirms that pollution reaches even the most remote areas of the ocean 2The Malaspina Expedition confirms that pollution reaches even the most remote areas of the ocean 3Unraveling cell division 2Unraveling cell division 3Cross Match Launches New Identity Management Handheld Solution 2
... ROCKVILLE, Md. -- Several members of the American Society ... or foreign associates of the U.S. National Academy of ... in original research. Three current ASPB members were ... Birchler , Curators Professor of Biological Sciences, University of ...
... Callaghan, a researcher at the Royal Swedish Academy of Sciences, ... and permafrost. "The changes we see are dramatic. ... deviate from the norm when compared with a longer term ... parts of the globe that is warming up fastest today. ...
... and Shinya Yamanaka, MD, PhD, have been elected as members ... in original scientific research. It is one of the highest ... States. They join 70 other scientists who will ... now has 40 current faculty members in the distinguished group. ...
Cached Biology News:A new research report shows effects of climate change in the Arctic are more extensive than expected 2A new research report shows effects of climate change in the Arctic are more extensive than expected 3UCSF scientists honored by National Academy of Sciences 2