OVERVIEW
- PCR is used to amplify a specific region of a DNA strand (DNA target). Most PCR methods typically amplify DNA fragments of up to ~10kb (our case). 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.
-In the PCR experiment, the experiment begins with the preparation of the master mix of the above mentioned components. The second step in the experiment is adding a specific amount of the master mix to the DNA template that contains the DNA region (target) to be amplified that you previously prepared in the DNA extraction lab.
-After adding the master mix to the DNA template, the tube will be placed in the PCR device and the device will be set so that the DNA sample will pass by four stages inside the PCR device. These stages are:
- Stage one: Initialization denaturation.
- Stage two: Multiplication cycles.
- Stage three: Final elongation.
- Stage four: Final hold.