Pcr (polymerase chain reaction) is a commonly used method for the amplification of a short segments of dna. the flow chart below
is a simplified illustration of the basic steps of pcr. complete the flow chart. if you answer any part of this question incorrectly, a single red x will appear indicating that one or more of the phrases are sorted incorrectly.
Extraction of DNA, Denaturation, Annealing, Polymerization.
Explanation:
Polymerase chain reaction (PCR) is a routine laboratory technique used to make many copies of a specific region of DNA. The purpose of PCR is to fabricate enough of the DNA region of interest so that it can be analyzed and used in some other way.
Extraction: The first step is to get the genetic material to be used, which in this case is DNA. It cannot be altered or contaminated by any other reagent that causes a different result than expected from the final combination. This extraction will be heated to the point of being denatured along with the region that needs to be amplified.
Denaturation is divided into two parts. For this step of the PCR technique to work, the handler must add a nitrogenous base composed of a three phosphate, the oligonucleotides (known as DNA strands A, T, C and G) and the polymerase to be used, to promote the reaction. Mixing should be done so that the DNA can conform to consistency and properly absorb the genetic information from the other three materials. Then the handler must put the mixture into a device called a thermal cycler. This device will promote specific temperature variations in the buffer solution containing the DNA and the mixture. This temperature instability helps turn DNA strands into a single structure, denatured. The heating comes first, with a maximum temperature of 95 ° for the strips to separate. Then comes the cooling, with the thermal cycler having a temperature of 50 °;
Annealing is the phase that will join the DNA strands (primers) on each complementary side. From there, the work of amplification begins, as the mold of the new molecule grows because of the combination of these complements. The annealing occurs during cooling in the thermal cycler and, after this period, the device warms up again so that the polymerase acts in the composition;
Polymerization: is the ultimate extension of DNA. In the PCR technique, this phase already presents the polymerase adding the nitrogenous bases. For this extension to gain strength and to be able to reproduce a new molecule, the mixture must be heated to a maximum temperature of 72 °.
