qPCR

Question 1

How to evaluate PCR

Answer 1

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Question 2

What is melting curve, CT- value

Answer 2

melting curve- Melting curve analysis is an assessment of the dissociation characteristics of double-stranded DNA during heating. As the temperature is raised, the double strand begins to dissociate leading to a rise in the absorbance intensity, hyperchromicity.

Ct value is the cycle number when the fluorescence of a PCR product can be detected above the background signal and corresponds to the number of cycles to reach a defined threshold.
Delta Ct corresponds to the difference between Ct of your gene of interest and Ctof your reference or housekeeping gene.

Question 3

common reason for bad qPCR

Answer 3

Poor quality RNA.
Selecting the detection chemistry.
Not using a master mix and organizing your experiments.
Skimping on controls.
Not validating your reference gene.

Question 4

what is PCR

Answer 4

Polymerase chain reaction is used to make millions of copies of target proteins

Question 5

What is qPCR or Real-Time RT-PCR?

Answer 5

is used to assay gene expression by measuring cDNA targets when RNA is used as the starting material.