Question

With your fragment now in an expression plasmid, you will need to check to ensure that the fragment is in the correct reading frame from the promoter so that a functional enzyme can be made. You will utilize the polymerase chain reaction (PCR) to amplify a section of the plasmid containing your insert so that it can be sequenced. PCR uses a DNA polymerase isolated from a hyperthermophilic bacterial species to synthesize a new DNA strand off a template strand of DNA. You are supplying a template in the form of the plasmid with the inserted fragment. PCR also uses the requirement of DNA polymerase to have a pre-existing primer with an open 3' end for new strand synthesis. You will provide the primers that will bind to specific DNA sequences before and after the inserted fragment and will act as boundaries for what part of the plasmid will be amplified. The resulting short, amplified fragments will be identical in base sequence to the original plasmid templates. Once PCR amplification is complete, you will have enough template fragments to send for DNA sequencing to ensure that you have inserted the genes correctly into the plasmid. Correct insertion will result in the production of an mRNA strand that will have the first gene codon in the correct reading frame for a working enzyme to be produced. Of the following components, which ones are required for the amplification of a specific section of DNA using the polymerase chain reaction? Select all that apply.
a. DNA primers
b. RNA polymerase
c. a strand of RNA to act as a template
d. DNA polymerase that is heat stable

Answer 1

Answer:

The correct answer is - a. DNA primers, and d. DNA polymerase that is heat stable.

Explanation:

In Polymerase chain reaction (PCR) there are many components are required in a overall process but there is two major components that are required for amplification of a specific section of DNA:

a. DNA primers- these are short and strands or sequence of ssDNA, a pair of these sequence acts as the starting site for a new strand which is used to hybridize with the sample DNA and mark the region of the DNA of interest to be amplified.

d. DNA polymerase that is heat stable-  PCR technique involves various reaction that are performed at different temperatures. A thermophilic bacteria Thermus aquaticus is used to get this heat stable enzyme, Which lives in hot springs.

Thus, the correct answer is - a. DNA primers, and d. DNA polymerase that is heat stable.