A simple, accurate and universal method for quantification of PCR

作者：Boulter Nicky^*; Suarez Francia Garces; Schibeci Stephen; Sunderland Trevor; Tolhurst Ornella; Hunter Tegan; Hodge George; Handelsman David; Simanainen Ulla; Hendriks Edward; Duggan Karen

来源：BMC Biotechnology, 2016, 16(1): 27.

DOI：10.1186/s12896-016-0256-y

• qPCR
• Absolute quantification
• Universal calibrator
• Gene expression

Background: Research into gene expression enables scientists to decipher the complex regulatory networks that control fundamental biological processes. Quantitative real-time PCR (qPCR) is a powerful and ubiquitous method for interrogation of gene expression. Accurate quantification is essential for correct interpretation of qPCR data. However, conventional relative and absolute quantification methodologies often give erroneous results or are laborious to perform.
To overcome these failings, we developed an accurate, simple to use, universal calibrator, AccuCal.
Results: Herein, we show that AccuCal quantification can be used with either dye-or probe-based detection methods and is accurate over a dynamic range of >= 10(5) copies, for amplicons up to 500 base pairs (bp). By providing absolute quantification of all genes of interest, AccuCal exposes, and circumvents, the well-known biases of qPCR, thus allowing objective experimental conclusions to be drawn.
Conclusion: We propose that AccuCal supersedes the traditional quantification methods of PCR.

• 出版日期2016-3-9