What is the role of dNTPs in PCR?

Deoxynucleoside triphosphates (dNTPs) dNTPs consist of four basic nucleotides—dATP, dCTP, dGTP, and dTTP—as building blocks of new DNA strands. However, dNTPs exceeding optimal concentrations can inhibit PCR.
A.

What does magnesium do in PCR?

Magnesium is required as a co-factor for thermostable DNA polymerase. Taq polymerase is a magnesium-dependent enzyme and determining the optimum concentration to use is critical to the success of the PCR reaction.
  • What is the role of Taq polymerase in PCR?

    T. aquaticus is a bacterium that lives in hot springs and hydrothermal vents, and Taq polymerase was identified as an enzyme able to withstand the protein-denaturing conditions (high temperature) required during PCR. Therefore, it replaced the DNA polymerase from E. coli originally used in PCR.
  • How many copies of DNA should be present after 30 cycles of PCR?

    The result, after 30 cycles of amplification, is the production of {230 - (2 x 30)} (ie. over one hundred million copies) of the DNA sequence in between, and including, the two primers.
  • What are four important applications of PCR?

    The polymerase chain reaction has been elaborated in many ways since its introduction and is now commonly used for a wide variety of applications including genotyping, cloning, mutation detection, sequencing, microarrays, forensics, and paternity testing. Typically, a PCR is a three-step reaction.
B.

Why do you need buffer in PCR?

Isopropanol precipitation of DNA and treatment of DNA pellets with 70% ethanol is usually effective in removing traces of contaminants from the DNA sample. PCR buffer is necessary to create optimal conditions for activity of Taq DNA polymerase. Buffers often contain Tris-Hcl, KCl, and sometimes MgCl2.
  • Why DNTP is used in PCR?

    The purpose of the deoxynucleotide triphosphates (dNTPs) is to supply the “bricks.” Since the idea behind PCR is to synthesize a virtually unlimited amount of a specific stretch of double-stranded DNA, the individual DNA bases must be supplied to the polymerase enzyme.
  • What is the role of Taq polymerase in PCR?

    T. aquaticus is a bacterium that lives in hot springs and hydrothermal vents, and Taq polymerase was identified as an enzyme able to withstand the protein-denaturing conditions (high temperature) required during PCR. Therefore, it replaced the DNA polymerase from E. coli originally used in PCR.
  • What is the function of the DNA polymerase in PCR?

    The most important enzyme in a PCR reaction is called taq polymerase. A polymerase is an enzyme that attaches molecules together, and we just so happen to want to attach many nucleotides (the building blocks of DNA) together, so it works out for us.

Updated: 2nd October 2019

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