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. View Available Hint (s) Select all that apply.

A. a strand of RNA to act as a template

B. DNA primers

C. RNA polymerase

D. DNA polymerase that is heat stable

B. DNA primers

D. DNA polymerase that is heat stable

Explanation:

The technique of DNA strand multiplication is called PCR (polymerase chain reaction). In the 1980s, the PCR technique was used to make thousands of copies of a single piece of DNA. This technique is used in test tubes containing the DNA and some needed compounds, such as primers (DNA primers) and the DNA polymerase enzyme (DNA replication enzyme).

Primers are DNA strands, with about 20 bases (A, T, C, G) complementary, that is, they bind by complementarity to the beginning of the DNA sequence to be multiplied. When a DNA molecule is to be multiplied, the double strand must be separated, thus forming two different but complementary strands. Each ribbon will serve as a template for duplication, so we need two different types of primers.

The polymerase chain reaction is as follows:

The DNA sample, the replication enzyme (DNA polymerase), the DNA nucleotides, and the DNA sequence complementary primers are placed in a test tube. The test tube is placed in a PCR machine (a machine that raises and lowers the temperature according to a program). The following heating and cooling steps take place inside the machine controlled by the program. The tube is heated to 94 ° C to denature (separate the double strand) the DNA. Due to the high temperature, the DNA polymerase must be heat stable. Each single strand of denatured DNA serves as a template for the synthesis of new complementary strands. For this it is cooled to 54ºC where the primers ring to the beginning of the two simple strips, serving as primers for the polymerase enzyme. The tube is reheated to 72 ° C (ideal DNA polymerase operating temperature) for the duplication of the strip. DNA polymerase begins, after the end of the primer, to place the free nucleotides on the DNA strand by binding them together, thus forming a new double strand