- The polymerase chain reaction (PCR) is one of the most powerful technologies in molecular biology. In conventional PCR, detection and quantification of the amplified sequence are performed at the end of the reaction after the last PCR cycle, and involve post-PCR analysis such
as gel electrophoresis and image analysis. In real-time PCR, PCR product is measured at each cycle.

The advantages of real-time PCR include:

  • Ability to monitor the progress of the PCR reaction as it occurs in real time.
  • Ability to precisely measure the amount of amplicon at each cycle, which allows highly accurate quantification of the amount of starting material in samples.
  • An increased dynamic range of detection.
  • Amplification and detection occurs in a single tube, eliminating post-PCR manipulations.
- In the real time PCR, we get threshold cycle which is the cycle number at which the fluorescent signal of the reaction crosses the threshold then through the standard curve the concentration of each sample could be calculated.
- The standard curve is a dilution series of known template concentrations can be used to establish a standard curve for determining the initial starting amount of the target template in experimental samples.
- A PCR setup requires several components and reagents. These components include:
  • Two primers (Forward and reverse).
  • Taq polymerase.
  • Deoxynucleoside triphosphates (dNTPs).
  • Buffer solution.
  • Distilled water.
  • Template DNA.
  • 2x SYBR Green


  • To amplify a targeted DNA molecule during the PCR in real-time.


  • Real-time polymerase chain reaction.

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