Iowa State University
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News Service

News Service:

Annette Hacker, director,
(515) 294-3720

Office: (515) 294-4777

03-09-07

Contacts:

Jack Gallup, Veterinary Pathology, (515) 294-5844, eag@iastate.edu

Teddi Barron, News Service, (515) 294-4778, tbarron@iastate.edu

Iowa State researcher develops software that sidesteps effects of qPCR-inhibitory materials and ensures precise and swift set-ups

AMES, Iowa - Although qPCR (fluorogenic real-time quantitative polymerase chain reaction) is considered one of the most important tools in contemporary molecular biology, its proper execution has become controversial in recent years. A widely shared view is that contaminants in samples tested by qPCR have skewed many research results dramatically by inhibiting the two main enzymatic reactions underlying PCR.

An Iowa State University researcher has developed a Microsoft Excel-based system that provides a way to detect and avoid inhibition, and enables investigators to consistently design dynamically sound, truly LOG-linear qPCR reactions. PREXCEL-Q was created by Jack Gallup, assistant scientist in veterinary pathology. The description of the invention was published in the Sept. 15, 2006 issue of Biological Procedures On-line.

PREXCEL-Q circumvents the effect of qPCR-inhibitory material in all qPCR assays by exactly calculating the safe (non-inhibitory) dilutions for all samples and standards (without over-diluting any of them) thereby allowing the assay to achieve its hallmark desired LOG-linear amplifications that are truly reflective of the absolute and/or relative abundance of the nucleic acid targets of interest in the samples. It does so by first defining the legitimate boundaries of each qPCR assay by examining representative mixtures of all samples in each assay beforehand.

The information gathered from this preliminary test allows the extent of qPCR-inhibitory behavior introduced by the samples themselves to be precisely measured and then subsequently avoided by strategic dilutions of each sample preceding final qPCR evaluation. The program's consistency, reliability, repeatability, precision and accuracy have been demonstrated in practice numerous times in the lab. It has been used at Iowa State University for the past seven years in Dr. Mark Ackermann's laboratory, and is being beta-tested by several Iowa State researchers.

PREXCEL-Q allows scientists to unveil the absolute or relative presence of any stretch of genetic material of interest in any biological sample as quickly and precisely as possible. The new program swiftly and accurately calculates all set-up parameters for all types of qPCR set-ups, and ensures that the gene expression analyses ultimately accomplished by the technique are unquestionably precise. PREXCEL-Q solves problems universal to all qPCR reactions, and performs all necessary qPCR set-up calculations in less than 15 minutes for up to seven qPCR targets and 72 samples at a time-calculations that might take days to complete otherwise.

PREXCEL-Q is available to academic institutions at no cost. Contact Jack Gallup, (515) 294-5844 or eag@iastate.edu.

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Quick look

"PREXCEL-Q is available to academic institutions at no cost. Contact Jack Gallup, (515) 294-5844 or eag@iastate.edu