qPCR and dPCR: Quantitation Methods in Molecular Biology

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Questions and Answers

What is the primary purpose of using a reference gene in qPCR analysis?

To amplify the target gene sequence
To identify the gene of interest in a sample
To quantify the absolute number of gene copies
To normalize the gene expression levels across different samples (correct)

What is the relationship between the fluorescence intensity and the amount of amplicon produced in a qPCR reaction?

Fluorescence intensity increases proportionally with the amount of amplicon produced (correct)
Fluorescence intensity decreases as the amount of amplicon produced increases
Fluorescence intensity remains constant regardless of the amount of amplicon produced
Fluorescence intensity is inversely proportional to the amount of amplicon produced

What is the formula to calculate the fold change in gene expression using the ΔΔCt method?

ΔΔCt × 2
2^(ΔΔCt) (correct)
2^(-ΔΔCt)
ΔΔCt / 2

What is the role of the cycle threshold (Ct) value in qPCR analysis?

To quantify the relative amount of amplicon produced (D) Signup and view all the answers

What is the purpose of creating a standard curve in qPCR analysis?

To determine the absolute number of gene copies (C) Signup and view all the answers

What is the effect of treatment on the expression of the gene TNF, as calculated by the ΔΔCt method?

A 32-fold increase in TNF expression (A) Signup and view all the answers

What is the difference between absolute and relative quantification in qPCR analysis?

Absolute quantification measures the absolute number of gene copies, while relative quantification measures the fold change (C) Signup and view all the answers