A Tale of Tails: Dissecting the Enhancing Effect of Tailed Primers in Real-Time PCR.

Last updated on 2-10-2019 by Anonymous (not verified)

Peer reviewed scientific article

SCIENSANO

Abstract:

Non-specific tail sequences are often added to the 5’-terminus of primers to improve the robustness and overall performance of diagnostic assays. Despite the widespread use of tailed primers, the underlying working mechanism is not well understood. To address this problem, we conducted a detailed in vitro and in silico analysis of the enhancing effect of primer tailing on 2 well-established foot-and-mouth disease virus (FMDV) RT-qPCR assays using an FMDV reference panel. Tailing of the panFMDV-5UTR primers mainly affected the shape of the amplification curves. Modelling of the raw fluoresce…

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