Introduction to Quantitative PCR (recording)

These flashcards cover key concepts and definitions related to quantitative PCR as discussed in the lecture.

What are the main components of a quantitative PCR (qPCR) mix?

Primers, template DNA, Taq polymerase, buffer, magnesium ions, and dNTPs.

What is the key difference between conventional PCR and quantitative PCR?

Quantitative PCR quantifies the amount of PCR product in real-time during the amplification process.

How is PCR product detected in quantitative PCR?

By measuring fluorescence that increases as the PCR product accumulates.

What role does dye play in quantitative PCR?

Dye binds to double-stranded DNA and fluoresces to indicate the amount of PCR product.

What is the purpose of the threshold cycle (Ct) in quantitative PCR?

It indicates the cycle number at which the fluorescence crosses a predetermined threshold, allowing quantification of the starting template amount.

What happens to the fluorescence signal when PCR reagents are consumed?

The fluorescence plateau occurs when key reagents become limiting, slowing down the amplification process.

What is a common non-specific dye used in quantitative PCR?

SYBR Green.

What is a molecular beacon in the context of quantitative PCR?

An oligonucleotide probe with a hairpin structure that fluoresces when bound to the target DNA, releasing FRET.

What is the function of Taq polymerase in the qPCR process?

It synthesizes new DNA strands and can process dNTPs during the amplification.

What does FRET stand for, and how is it used in quantitative PCR?

Fluorescence Resonance Energy Transfer; it occurs when two fluorescent dyes are close, quenching fluorescence until they are separated by probe binding to the target DNA.