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#Biochemistry #voets
In PCR, a DNA sample is separated into single strands and incubated with DNA polymerase, dNTPs, and two oligonucleotide primers whose sequences flank the DNA segment of interest. The primers direct the DNA polymerase to synthesize com- plementary strands of the target DNA (Fig. 3-27). Multiple cycles of this process, each doubling the amount of the target DNA, geometrically amplify the DNA starting from as little as a single gene copy. In each cycle, the two strands of the duplex DNA are separated by heating, then the reaction mixture is cooled to allow the primers to anneal to their complementary segments on the DNA. Next, the DNA polymerase directs the synthesis of the complementary strands. The use of a heat-stable DNA polymerase, such as Taq polymerase isolated from Thermus aquaticus, a bacterium that thrives at 75°C, eliminates the need to add fresh enzyme after each round of heating (heat inactivates most en- zymes). Hence, in the presence of sufficient quantities of primers and dNTPs, PCR is carried out simply by cyclically varying the temperature.
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owner: smelly_compost - (no access) - Voet's Fundamentals of Biochemistry 4th Edition.pdf, p101


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