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Polymerase Chain Reaction is not used in
Polymerase Chain Reaction is not used in
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The correct option is C separating DNA fragments of genetic material isolated from a bacterial cell
Polymerase Chain Reaction is a process used to amplify DNA segments and obtain numerous copies of DNA in a short time.
This involves the use of two sets of primers (forward primer and reverse primer which attach to the 3’ OH ends of the DNA strands oriented in 3’→5’ and 5’→3’ polarity, respectively) and a thermostable DNA polymerase (Taq polymerase).
There are 3 steps involved in this process:
- Denaturation: Separation of the two strands of double stranded DNA by application of heat at 96°C.
- Annealing: Hybridisation of two primers with specific sequences of each of the strands of the DNA. This process is carried out at 55-65°C.
- Extension: Taq DNA polymerase synthesises DNA by adding dNTPs to the free ends of the two primers - forward and reverse. The extension of DNA is carried out at 72°C. This helps in synthesizing complementary strands. This entire process of in vitro DNA replication is repeated multiple times to obtain multiple copies of a DNA fragment.
Thus, PCR can be used to detect traces of DNA and can be used to confirm the presence of a pathogen during early stages of infection. PCR is now being extensively used in the detection of the HIV in suspected AIDS cases and to detect mutations in suspected cancer patients.
Extraction of genomic DNA is done by dissolving the cell wall and cell membrane of the cell from which gene of interest is to be isolated. Separation of the isolated DNA fragments is done using Agarose gel electrophoresis.
The DNA fragments and size markers are loaded onto the gel well and current is passed through the gel. The DNA being negatively charged, moves towards the positive electrode during gel electrophoresis. The smaller the size of the molecule, the faster and farther it travels. This is how the fragments are separated.
Polymerase Chain Reaction is a process used to amplify DNA segments and obtain numerous copies of DNA in a short time.
This involves the use of two sets of primers (forward primer and reverse primer which attach to the 3’ OH ends of the DNA strands oriented in 3’→5’ and 5’→3’ polarity, respectively) and a thermostable DNA polymerase (Taq polymerase).
There are 3 steps involved in this process:
- Denaturation: Separation of the two strands of double stranded DNA by application of heat at 96°C.
- Annealing: Hybridisation of two primers with specific sequences of each of the strands of the DNA. This process is carried out at 55-65°C.
- Extension: Taq DNA polymerase synthesises DNA by adding dNTPs to the free ends of the two primers - forward and reverse. The extension of DNA is carried out at 72°C. This helps in synthesizing complementary strands. This entire process of in vitro DNA replication is repeated multiple times to obtain multiple copies of a DNA fragment.
Thus, PCR can be used to detect traces of DNA and can be used to confirm the presence of a pathogen during early stages of infection. PCR is now being extensively used in the detection of the HIV in suspected AIDS cases and to detect mutations in suspected cancer patients.
Extraction of genomic DNA is done by dissolving the cell wall and cell membrane of the cell from which gene of interest is to be isolated. Separation of the isolated DNA fragments is done using Agarose gel electrophoresis.
The DNA fragments and size markers are loaded onto the gel well and current is passed through the gel. The DNA being negatively charged, moves towards the positive electrode during gel electrophoresis. The smaller the size of the molecule, the faster and farther it travels. This is how the fragments are separated.
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