Will u explain me the 3 rd step involved in rDNAtechnology ie; amplification of gene of interest using PCR - Biology - Biotechnology Principles and Processes

Question

Will u explain me the 3 rd step involved in rDNAtechnology ie;
amplification of gene of interest using PCR ??

Mushtaq Ahmed · Accepted Answer

Polymerase Chain Reaction (PCR)

Recombinant DNA can be amplified by PCR Several identical copies
of it can be synthesised in vitro

Two sets of primers (chemically synthesised
oligonucleotide stretches that are complementary to a region of
DNA), enzyme DNA polymerase,and deoxynucleotides
are added

PCR consists of 3 steps:

Denaturation − Double helical DNA is
denatured by providing high temperature DNA polymerase does not get
degraded in such high temperatures since the DNA polymerase used in
this reaction is thermostable as it is isolated from thermophilic
bacteria, Thermus aquaticus (Taq)

Annealing- It is the step in which primers are
annealed to single stranded DNA templates Two sets of primers
(small chemically synthesised oligonucleotides that are
complementary to the regions of DNA) are used The temperature of
reaction mixture is lowered to 50- 65°C for some seconds to
allow annealing of primers DNA polymerase extends the primer in 5'
to 3' direction

Extension − Replication of DNA occurs in
vitro

This cycle is repeated several times to generate up to 1 billion
identical copies of the DNA

Will u explain me the 3 rd step involved in rDNAtechnology ie; amplification of gene of interest using PCR ??