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Multiplex PCR Optimization

DNA Software > Articles by: joseph@dnasoftware.com

Author: joseph@dnasoftware.com

Multiplex PCR Optimization

Posted on by joseph@dnasoftware.com

Myth 7: Multiplex PCR Can Succeed by Optimization of Individual PCRs

 

Not too many people believe this myth, and yet their actions are somewhat irrational as they proceed to immediately use that approach to try to experimentally optimize a multiplex

Read more ›

Amplification Efficiency for primers Is Not Exponential

Posted on by joseph@dnasoftware.com

Myth 6: At the End of PCR, Amplification Efficiency Is Not Exponential Because the Primers or NTPs Are Exhausted or the Polymerase Looses Activity

Exponential amplificatiion of DNAPCR amplification occurs with a characteristic “S” shape. During the early cycles of PCR, the

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Why BLAST Search is a Myth for Determining the Specificity of a Primer

Posted on by joseph@dnasoftware.com

Myth 5: A BLAST Search Is the Best Method for Determining the Specificity of a Primer

 

To minimize mispriming, several PCR texts suggest performing a BLAST search, and such capability is a part of some primer design packages such

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Primer Dimer Artifacts Are Due to Dimerization

Posted on by joseph@dnasoftware.com

Myth 4: “Primer Dimer” Artifacts Are Due to Dimerization of Primers

 

A common artifact in PCR is the amplification of “primer dimers.” The most common conception of the origin of primer dimers is that two primers hybridize at their 3′-ends

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Designing Forward and Reverse Primers to Have Matching Tm

Posted on by joseph@dnasoftware.com

DNA helical structure

Myth 3: Designing Forward and Reverse Primers to Have Matching Tm’s Is the Best Strategy to Optimize for PCR

Nearly all “experts” in PCR design would claim to believe in myth 3. Most current software packages base their design strategy

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Best Methods for Predicting Hybridization Tm

Posted on by joseph@dnasoftware.com

Myth 2: Best Methods for Predicting Hybridization Tm

Best methods for predicting hybridization Tm are essentially equivalent in accuracy. The melting temperature, Tm, of duplex formation is usually defined as the temperature at which half the available strands are in

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PCR Working and Design Is Not Important

Posted on by joseph@dnasoftware.com

Myth 1: PCR Nearly Always Works and Design Is Not that Important

It might come as a surprise to many that despite the wide use and large investment, PCR in fact is still subject to many artifacts and environmental factors Read more ›

Comparison of traditional quantitative pcr with computer quantitation

Posted on by joseph@dnasoftware.com

Comparison of traditional quantitative pcr with a computer-based quantitation algorithm for cmv from plasma specimens

Background

Difference between traditional quantitative pcr with computer-based quantitation algorithm for cmv. Traditional viral quantitation using cycle threshold (Ct) analysis is dependent on generating an

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DNA Software releases ThermoBLAST Cloud Edition

Posted on by joseph@dnasoftware.com

DNA Software introduced the full commercial release of ThermoBLAST Cloud Edition (TB-CE). TB-CE provides a new standard for evaluating the target specificity of oligonucleotides.

Click here to learn more

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DNAS gives webinar on qPCR CopyCount

Posted on by joseph@dnasoftware.com

DNAS gives webinar on qPCR CopyCount, click here to view video.

 

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