Full Form : Reverse transcription polymerase chain reaction
Definition:-
- It is a laboratory technique combining reverse transcription of RNA into DNA and amplification of specific DNA target using PCR.
- It is primarily used to measure the amount of a specific RNA.
- RT-PCR is a nuclear-derived method for detecting the presence of specific genetic material in any pathogen, including a virus.
Principles:-
One-step RT-PCR vs two-step RT-PCR
- The quantification of mRNA using RT-PCR can be achieved as either a one-step or a two-step reaction.
- The difference between the two approaches lies in the number of tubes used when performing the procedure.
- The two-step reaction requires that the reverse transcriptase reaction and PCR amplification be performed in separate tubes.
4. The disadvantage of the two-step approach is susceptibility to contamination due to more frequent sample handling.
End-point RT-PCR vs real-time RT-PCR
Application:-
- Research methods
- Gene insertion
- Genetic disease diagnosis
- Cancer detection
Advantages :-
- More accurate and less prone to combination
- Rapid
- Cheap
- Less error possibilities
Disadvantages:-
- Too difficult to be widely used
- Require expensive laboratory equipments
- Highly trained laboratory staff is needed
- Increase the risk of false negative or positive results
-Madiha Fatma ( MBBS & Academic Director, Editor – MCD )
