describe the provess of amplification of gene of interested using PCR techniques? draw diagram and name it's steps
Answers
Answer:
The amplification of gene is done by using the technique of PCR. PCR stands for Polymerase Chain reaction. PCR enables the production or amplification of billions of copies of an original piece of DNA in the tube with minutes or hours. A single PCR reaction involves three temperature-dependent steps, described as follow:
Denaturation: The starting solution is heated; usually to 94 degree Celsius. The high temperature breaks the hydrogen bonds between the two strands to the original DNA double helix, providing the necessary single-standard templates.
Annealing: After just a couple of minutes at that temperature, the reaction mixture is quickly cooled, usually to somewhere between 50 degree Celsius and 60 degree Celsius. It is then held for less than a minutes at this lower temperature- which is enough time for the primers to bind to their complementary sequences on the single-standard templates.
Primer extension (polymerization): The sample is next heated to 72 degree Celsius for some time, during which time the DNA polymerase adds nucleotides to the primer, synthesizing a new DNA strand using only the template sequences that bind the primers.
If the process of DNA replication is repeated many times, the segment of DNA can be amplified to approximately billion times, i.e., 1 billion copies are made at the end of 30 PCR cycles. It is possible to generate 2
n
molecules after 'n' number of cycles.
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Answer:
The amplification of gene is done by using the technique of PCR. PCR stands for Polymerase Chain reaction. PCR enables the production or amplification of billions of copies of an original piece of DNA in the tube with minutes or hours. A single PCR reaction involves three temperature-dependent steps, described as follow:
Denaturation: The starting solution is heated; usually to 94 degree Celsius. The high temperature breaks the hydrogen bonds between the two strands to the original DNA double helix, providing the necessary single-standard templates.
Annealing: After just a couple of minutes at that temperature, the reaction mixture is quickly cooled, usually to somewhere between 50 degree Celsius and 60 degree Celsius. It is then held for less than a minutes at this lower temperature- which is enough time for the primers to bind to their complementary sequences on the single-standard templates.
Primer extension (polymerization): The sample is next heated to 72 degree Celsius for some time, during which time the DNA polymerase adds nucleotides to the primer, synthesizing a new DNA strand using only the template sequences that bind the primers.
Explanation: