English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Plus annual

Presents the access of premium content as premium feature

Premium Content

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Plus monthly

Global: Zendy Tools annual

Global: Zendy Tools monthly

Global: Zendy Free Plan

Real-time PCR, or Real-time Quantitative PCR (qPCR) is an effective approachto quantify nucleic acid samples. Given the complicated reaction system alongwith thermal cycles, there has been long-term confusion on accuratelycalculating the initial nucleic acid amounts from the fluorescence signals.Although many improved algorithms had been proposed, the classical thresholdmethod is still the primary choice in the routine application. In this study,we will first illustrate the origin of the linear relationship between thethreshold value and logarithm of the initial nucleic acid amount byreconstructing the PCR reaction process with stochastic simulations. We thendevelop a new method for the absolute quantification of nucleic acid sampleswith qPCR. By monitoring the fluorescence signal changes in every stage of thethermal cycle, we are able to calculate a representation of the step-wiseefficiency change. This is the first work calculated PCR efficiency changedirectly from the fluorescence signal, without fitting or sophisticatedanalysis. Our results revealed that the efficiency change during the PCRprocess is complicated and can not be modeled simply by monotone functionmodel. Based on the calculated efficiency, we illustrate a new absolute qPCRanalysis method for accurately determining nucleic acid amount. The efficiencyproblem is completely avoided in this new method.

Absolute quantification of real-time PCR data with stage signal difference analysis