Demonstration of PCR and Gel Electrophoresis
Purpose
In this demo, we take a specific segment of Zebrafish DNA and multiply it by a billion. We then look at it under UV light
PCR (Polymerase Chain Reaction)
This tube contains
- Zebrafish DNA
- Primers (Primers attach themselves to beginning of the DNA segment we want to copy. The primers you choose determines what segment of DNA gets copied)
- Nucleotides (These get assembled to create the copies)
- Taq polymerase (Assembles the copy of the selected DNA segment )
- Magnesium Chloride
- Blue and yellow dyes (we’ll see these as separate color spots when we’re finished. The DNA that gets copied will be found between them)
To double the amount of the selected DNA segment, the temperature of the sample is changed to 94C, 55C then 72C. After doing this 30 times, we end up with about a billion times as much. The thermocycler handles the temperature changes.
The program
Gel Electrophoresis
Takes the DNA segments that were just created, separates them from the rest of the tube contents, then binds it with a florescent dye so we can see it under UV light.
Examining the results
Lane 1: A “ladder”.
Lane 2: Sample with DNA
Lane 3: Sample without DNA