Guidelines for validation of qualitative real-time PCR methods

Guidelines for validation of qualitative real-time PCR methods

S. Broeders*, I. Huber, L. Grohmann, G. Berben, I. Taverniers, M. Mazzara, N. Roosens and D. Morisset Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050 Brussels, Belgium (Tel.: D32 2 6425425; fax: D32 2 6425292; e-mails: [email protected]; nancy.roosens@ wiv-isp.be) Bavarian Health and Food Safety Authority (LGL), Veterin€arstraße 2, 85764 Oberschleißheim, Germ...

Robust regression methods for real-time PCR

Validation of Reference Genes for Real Time PCR Normalization in Milk Somatic Cells of Holstein Dairy Cattle

Real time-qPCR is the most reliable method for evaluation of mRNA expression levels. However, to obtain accurate results, selection of suitable reference genes is necessary for normalizing the real-time qPCR data. The aim of this research was to validate the expression stability of three potential reference genes (ACTB, GAPDH and UXT) in milk somatic cells of Holstein dairy cattle under differe...

Checklist for optimization and validation of real-time PCR assays.

Real-time polymerase chain reaction (PCR) is a frequently used technique in molecular diagnostics. To date, practical guidelines for the complete process of optimization and validation of commercial and in-house developed molecular diagnostic methods are scare. Therefore, we propose a practical guiding principle for the optimization and validation of real-time PCR assays. Based on literature, e...

Increased efficacy for in-house validation of real-time PCR GMO detection methods

To improve the efficacy of the in-house validation of GMO detection methods (DNA isolation and real-time PCR, polymerase chain reaction), a study was performed to gain insight in the contribution of the different steps of the GMO detection method to the repeatability and in-house reproducibility. In the present study, 19 methods for (GM) soy, maize canola and potato were validated in-house of w...