qPCR Flashcards
1
Q
What does qPCR stand for?
A
Real-Time or Quantitative PCR
2
Q
What is purpose of qPCR?
A
Amplification of DNA by polymerase chain reaction (PCR)
3
Q
What does Probe have?
A
5’ fluorescent label (6-FAM), 3’ quencher (BHQ)
4
Q
Where does probe attach to?
A
In the DNA template between the FWD and REV primers
5
Q
What activity does DNA polymerase have?
A
5’ to 3’ exonuclease activity
6
Q
What part of the probe does DNA polymerase cleave?
A
5’ fluorescent label of probe and signal intensifies when separated from quencher
7
Q
How is DNA amplification detected?
A
fluorescence is detected when fluorescence tag and quencher are separated
8
Q
What kind of quantification does qPCR have?
A
relative quantification, uses standard curve to calculate amount of sample amplified
