Real time PCR

Question 1

What is real time PCR

Answer 1

Real time PCR is a type of PCR that doesn’t need you to use gel electrophoresis to view the bands after.

Question 2

How is real time PCR powerful

Answer 2

it quantifies gene expression which is a particular gene that is being expressed in a cell and to what level it is being expressed

Question 3

How does standard PCR amplify the target region?

Answer 3

Standard PCR doubles the amount of amplicon with each cycle, amplifying exponentially until reagents are depleted, typically around 30 cycles.

Question 4

How does qPCR address the limitation of standard PCR?

Answer 4

qPCR allows for a direct correlation between the starting amount of target in a sample and the amount of target produced at the end of PCR.

Question 5

What phase of the PCR cycle does qPCR primarily focus on for comparison?

Answer 5

qPCR primarily focuses on the early exponential phase for comparison between samples.

Question 6

Why can’t standard PCR detect amplification during the early exponential phase?

Answer 6

Standard PCR cannot detect amplification during the early exponential phase because the amount of amplicon produced at that point is too low to be visible on a gel.

Question 7

What does qPCR rely on for accurate detection during the early exponential phase?

Answer 7

qPCR relies on very sensitive detection instrumentation to accurately determine the level of fluorescence emitted from a sample during PCR.

Question 8

what are the two types of fluorescence systems that can be used

Answer 8

SYBR green and TaqMan probe