describe this method Reverse transcriptionPCR How and when w

describe this method:

Reverse transcription-PCR

How and when would you use these two methods? How are they helpful in molecular biology?

Solution

Polymerase chain reaction (PCR) is a biotechnological method to replicate DNA (deoxyribonucleic acid) in the laboratory, and multiple copies of the same sequence can be produced. The sequence of steps in PCR is as follows.

The unwinding of DNA strands is done by slight heating.

Artificially synthesised primer and DNA polymerase are added along with the four deoxyribonucleotide triphosphates namely, dATP, dGTP, dCTP, dTTP. These supply the four bases namely, adenine (A), thymine (T), guanine (G), and cytosine (C), which are the building blocks of DNA’s double helical structure.

The elongation of complementary strand is catalysed by DNA polymerase.

Reverse transcription polymerase chain reaction (RT-PCR) is a variant of traditional PCR technique, which is used to detect RNA expression through the creation of complementary DNA (cDNA) transcripts from RNA.

Both the traditional PCR and RT-PCR generate multiple copies of DNA fragments through gene amplification. The traditional PCR is used to amplify the specific DNA sequences, whereas the RT-PCR is used to amplify the genes by reverse transcription of RNA into its complementary DNA through reverse transcription technique. This complementary DNA strand is now amplified through traditional PCR.