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:10/20/2014)... , Oct. 20, 2014 Mapp ... experimental ZMapp™ antibody therapeutic to fight the Ebola ... difficult and time-consuming the production of pharmaceuticals can ... research publisher said that while some may be ... this compound, those with industry knowledge are well ...
(Date:10/20/2014)... , October 20, 2014 , ... (GI) disorders and the delivery of care has revealed ... and worrying inequalities in the provision of healthcare services ... was commissioned by United European Gastroenterology (UEG), have been ... and public awareness of the burden of GI disorders ...
(Date:10/20/2014)... Virginia (PRWEB) October 20, 2014 ... NDA Partners LLC, announced today that Ellen Teplitzky, ... legal support for the pharmaceutical industry, has joined ... manager of its legal services practice. NDA ... and expert witness and testimony, to top law ...
(Date:10/19/2014)... 2014 NextCODE Health, which enables clinicians and researchers ... time, today announced the launch of its new genomic data ... the American Society of Human Genetics (ASHG) annual meeting in ... up and apply for free beta access, visit www.nextcode.com ... can be viewed here . The Exchange: ...
Breaking Biology Technology:Kalorama: ZMapp Highlights Need For Faster Biopharmaceutical Production 2Kalorama: ZMapp Highlights Need For Faster Biopharmaceutical Production 3Major Survey Reveals Changing Trends and Inequalities in Healthcare Provision for Gastrointestinal Disorders Across Europe 2Major Survey Reveals Changing Trends and Inequalities in Healthcare Provision for Gastrointestinal Disorders Across Europe 3Major Survey Reveals Changing Trends and Inequalities in Healthcare Provision for Gastrointestinal Disorders Across Europe 4Major Survey Reveals Changing Trends and Inequalities in Healthcare Provision for Gastrointestinal Disorders Across Europe 5NDA Partners Appoints Ellen Teplitzky, JD as Director of its Legal Services Practice 2The NextCODE Exchange: The first global, real-time system for sharing full-resolution genomic data 2The NextCODE Exchange: The first global, real-time system for sharing full-resolution genomic data 3
... SAN DIEGO, Nov. 2 Cardium Therapeutics (NYSE Amex: ... 510(k) premarket notification with the U.S. Food and Drug ... product candidate. Excellagen XL is an ... customized collagen-based topical gel designed for use by physicians ...
... and LONDON, Nov. 2 A study from Thomson Reuters ... far outpacing research activity in the rest of the world. ... within the next decade. , The study, Global ... collaboration potential of China and its current place in world ...
... Pharmasset, Inc. (Nasdaq: VRUS ) today announced that it ... the Study of Liver Diseases (AASLD) on Monday, November 2, ... review Pharmasset,s progress on the programs that are the subject ... of this event via the internet, log on to the ...
Cached Biology Technology:Cardium Announces Plans to File FDA 510(k) Application for U.S. Marketing Clearance of ExcellagenXL(TM) Gel for Diabetic, Pressure and Venous Ulcers and Other Topical Wounds 2Cardium Announces Plans to File FDA 510(k) Application for U.S. Marketing Clearance of ExcellagenXL(TM) Gel for Diabetic, Pressure and Venous Ulcers and Other Topical Wounds 3Cardium Announces Plans to File FDA 510(k) Application for U.S. Marketing Clearance of ExcellagenXL(TM) Gel for Diabetic, Pressure and Venous Ulcers and Other Topical Wounds 4China's Research Output More Than Doubled Since 2004, Thomson Reuters Study Reveals 2Pharmasset to Webcast an Investor Event from the AASLD Meeting 2Pharmasset to Webcast an Investor Event from the AASLD Meeting 3Pharmasset to Webcast an Investor Event from the AASLD Meeting 4
(Date:10/16/2014)... – Post-menopausal women experienced improvements in vaginal atrophy, ... genital bleeding, after 12 weeks of daily 10 ... germ-based nutritional supplement previously shown to help relieve ... pilot study reported in a poster at the ... , "These data documented improved vaginal epithelium, ...
(Date:10/15/2014)... book ,Models of Life, is a non-traditional biophysics textbook ... The book is a journey of discovery into biological ... life and biological regulation. It is about how our ... communication between the billions of cells in an organism. ... relationships between principles, which can be found on both ...
(Date:10/15/2014)... NXT-ID, Inc. (NASDAQ: NXTD and NXTDW) ... growing mobile commerce market releases photos and video of the recent ... October 13 th . Gino Pereira , ... angel investor Mr. Chad A. Verdi rang the bell.  ... his investors and employees "for their work and dedication in bringing ...
Breaking Biology News(10 mins):S-equol supplements associated with improved measures of reproductive health in postmenopausal women 2New book about life as seen from physics 2Photo Release: NXT-ID Inc. Rings Opening Bell of the NASDAQ Stock Market October 13th 2Photo Release: NXT-ID Inc. Rings Opening Bell of the NASDAQ Stock Market October 13th 3
... time that several common species of seaweeds in both the ... chemical means. While competition between seaweed and ... of coral reefs worldwide, this chemical threat may provide a ... Seaweeds are normally kept in check by herbivorous fish, but ...
... hard winter for Monarch butterflies, according to Chip Taylor, director ... that low temperatures, storms and habitat destruction have all threatened ... a lot of time fretting over the status of the ... that will help me understand the past as a way ...
... from Rensselaer Polytechnic Institute will send an army of ... of preventing the formation and spread of biofilms, or ... the health of astronauts. The Micro-2 experiment, led ... Chemical and Biological Engineering at Rensselaer, is scheduled to ...
Cached Biology News:Researchers offer first proof that chemicals from seaweeds damage coral on contact 2Researchers offer first proof that chemicals from seaweeds damage coral on contact 3Researchers offer first proof that chemicals from seaweeds damage coral on contact 4As monarch butterflies journey north, gardeners can help protect species, researcher says 2Rensselaer researchers to send bacteria into orbit aboard space shuttle Atlantis 2
Umbilical cord blood neutrophils, For immunohistochemistry (IHC)...
...
Adjustable tilt angle 0 to 20 Adjustable speed of 3-25 RPM Load capacity: 2.7 kg (6 lbs.) Ambient temperature range 4C to 65C Low voltage power is supplied by wall mount transformer...
... Clean spin filters allow for the rapid isolation ... your DNA gel slice in the spin filter, ... your DNA is recovered. There is no need ... buffers. Spin filters can also be used for ...
Biology Products: