Genetics

qPCR in molecular diagnostics

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Explore the fundamentals behind qPCR data analysis. In a novel scenario, define and build a qPCR cycle plot, calculate cycle threshold, build and plot a standard curve and quantify an unknown sample. Use the data you collect to reach an evidence-based conclusion in context of the scenario.

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Learning outcomes

Interpret the different phases of a standard qPCR curve and deduce the start and end of each cycle.
Distinguish between the different signal traces for probes, reference and quencher and explain their purposes.
Construct and analyse a standard curve and interpret CT values.
Calculate gene expression data by extrapolation from a standard curve and describe the impacts of positive and negative ΔCT values.
Conclude whether a food product is safe for human consumption based on prerequisite evidence and gene expression data.

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Skills

Manipulating and Presenting Data

Defining the logarithmic plot parameters and defining the fluorescence threshold
Defining the axes and trendline of the standard curve
How to calculate the template concentration using the standard curve
How to quantify the ΔΔCT and FC for all the samples required

Interpreting and Analysing Data

Identifying the different phases of an amplification plot and determining the cycle start and end values
Identifying the differences between the probe, reference and quencher signals
Identifying a baseline and cycle threshold from an amplification plot
Identifying the CT value from the amplification plot
Concluding the meaning of a negative ΔCT
Assessing the nature of the fold difference observed and determining whether or not the food was safe for consumption

Concepts, Principles and Theories

Identifying the reasons why the reference signal is substracted from the probe signal

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Prof. Nicholas Harmer

University of Exeter

“We would like to bring Smart Worksheets into more areas because they're transformative in terms of getting students to really get to grips with problems and to upskill themselves numerically.”

Details

Degrees

Biological science

Biomedical science

Foundation science

Modules

Molecular biology

Topic

Genetics

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