"
Team:Evry/Protocols/07
From 2013.igem.org
| Line 16: | Line 16: | ||
<p>Polymerase Chain Reaction (PCR) is a molecular biology method used to amplify a small amount of genetic material (DNA or RNA), using specific primers of a target sequence. <br> | <p>Polymerase Chain Reaction (PCR) is a molecular biology method used to amplify a small amount of genetic material (DNA or RNA), using specific primers of a target sequence. <br> | ||
PCR is divided into 5 steps:<br> | PCR is divided into 5 steps:<br> | ||
| - | - First denaturation<br> | + | - First denaturation:<br> |
- Denaturation step<br> | - Denaturation step<br> | ||
- Annealing step<br> | - Annealing step<br> | ||
| Line 23: | Line 23: | ||
The 2nd, 3th and 4th steps are repeated 20-40 cycles.<br></p> | The 2nd, 3th and 4th steps are repeated 20-40 cycles.<br></p> | ||
| - | Denaturation step | + | <p>Denaturation step occure between 94 and 98°C.The heat breaks the hydrogen bonds and then causes the separation of the double strand of DNA into two single strands. <br> |
| + | Annealing step occure between 50 and 65 °C. Primers anneals to DNA simple strand by complementarity of the nitrogenized bases. <br> | ||
| + | Elongation step occure between 70 and 80°C, depending on the DNA polymerase used. The polymerase synthesizes a new DNA strand complementary to the DNA template</p> | ||
Revision as of 13:20, 26 August 2013
Polymerase Chain Reaction
Principle
Polymerase Chain Reaction (PCR) is a molecular biology method used to amplify a small amount of genetic material (DNA or RNA), using specific primers of a target sequence.
PCR is divided into 5 steps:
- First denaturation:
- Denaturation step
- Annealing step
- Elongation step
- Final step
The 2nd, 3th and 4th steps are repeated 20-40 cycles.
Denaturation step occure between 94 and 98°C.The heat breaks the hydrogen bonds and then causes the separation of the double strand of DNA into two single strands.
Annealing step occure between 50 and 65 °C. Primers anneals to DNA simple strand by complementarity of the nitrogenized bases.
Elongation step occure between 70 and 80°C, depending on the DNA polymerase used. The polymerase synthesizes a new DNA strand complementary to the DNA template
Preparation
Optimisation
Number of primer
Temperature
Gel electrophoresis analysis
Principle
Preparation
Analysis
