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Polymerase Chain Reaction (PCR)
The polymerase chain reaction, or simply PCR, is a method of amplifying DNA sequences. It provides us with a quick, accurate and efficient way of producing millions or even billions of copies of some target DNA segment. In order to carry out a successful polymerase chain reaction, we need (1) target DNA segment (2) DNA primers complementary to the flanking sequence of the target DNA fragment (3) heat-resistant DNA polymerase (4) all four types of deoxyribonucleoside triphosphates. A single polymerase chain reaction can be broken down into three stages - DNA strand separation, DNA primer hybridization and DNA synthesis. During strand separation, a solution containing all the ingredients listed above is heated to a temperature of 95 degree Celsius. This breaks the hydrogen bonds between the two stands of DNA, thereby separating the double stranded DNA molecule into the two individual single-strands. During the hybridization stage, the solution is heated to 54 degrees Celsius. This is a temperature at which the DNA primers begin to hybridize with the 3' end of the single-stranded DNA molecule. During the DNA synthesis stage, the temperature is increased to 72 degrees Celsius. This is the optimal temperature at which the heat-resistant DNA polymerase begins to elongate and synthesize the DNA strands. At the end of this stage, two copies of DNA molecules are produced. This cycle can be repeated as many times as we'd like to produce a total number of 2^n copies, where n is the number of cycles we carried out.
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