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Optimize Your qPCR and RT-qPCR Assays with Careful Planning and Design

Nadine earned a B.S. degree in Molecular Biology from the University of Wisconsin and a M.S. degree in Biotechnology and Biomedical Science from the University of Massachusetts – Boston.  After completing her degrees, Nadine worked with a team at the University of Wisconsin Hospital and Clinics, investigating molecular alterations associated with the progression of prostate cancer.

In 1997, Nadine joined the Genetic Analysis team at Promega Corporation, helping to launch reagent systems for applications in nucleic acid technologies.  Products developed include genotyping assays, nucleic acid purification methods, microarray reagents for gene expression analysis, and qPCR systems for DNA and RNA quantitation and detection.

  • Nadine Nassif

  • Senior Research Scientist

  • Original Webinar Date: Tuesday, January 15, 2013

Real-time PCR (qPCR) has become the technique of choice for a variety of applications such a gene expression analysis, genotyping, specific target sequence detection/quantitation and overall nucleic acid quantitation. This webinar will cover multiple qPCR topics aimed at improving your qPCR results including: optimizing qPCR assays including whether to use probe- or dye-based detection, choosing between 1-step and 2-step RT-qPCR assays, and overcoming key challenges such as sample integrity, multiplexing and the presence of inhibitors in the input sample. We will also introduce the GoTaq® family of qPCR and RT-qPCR systems which help researchers overcome these challenges.

Information

Traditional (endpoint) PCR assays measure the amount of product produced after cycling is completed making it difficult to quantitate the amount of starting nucleic acid template. In this case, the number of cycles must be carefully chosen for each target such that amplification is within the logarithmic phase in order to get accurate quantitation. Real-Time PCR (quantitative PCR, qPCR) overcomes this problem by monitoring product accumulation as PCR amplification proceeds by measuring the change in a fluorescent signal that reports on amplicon abundance. This approach allows for higher sensitivity and more accurate quantitation. Since its inception, qPCR has become a standard tool in the analysis of nucleic acids for many applications such a gene expression analysis, genotyping, specific target sequence detection/quantitation and overall nucleic acid quantitation. 

This webinar will cover important topics including (i) optimizing qPCR assays including whether to use probe- or dye-based detection, (ii) choosing between 1-step and 2-step RT-qPCR assays, and (iii) overcoming key challenges such as sample integrity, multiplexing and the presence of inhibitors in the input sample. We will also introduce the GoTaq® family of qPCR and RT-qPCR systems which help researchers overcome these challenges.