PCR & Gel Electrophoresis

Amplify DNA sequences and separate fragments by size.

Protocol

Main Goal

To understand the principles of DNA amplification via PCR and the separation of DNA fragments by size using agarose gel electrophoresis.

Experimental Protocol

  1. 1

    Prepare the PCR reaction mix (Template, Primers, dNTPs, Taq Polymerase).

  2. 2

    Set the thermal cycler to run through 20-30 cycles of Denaturation, Annealing, and Extension.

  3. 3

    Observe the exponential increase in DNA copies on the Results graph.

  4. 4

    Load the amplified DNA sample into a well of the agarose gel.

  5. 5

    Load a DNA ladder (reference standards) into another well.

  6. 6

    Apply an electric field (100V) and allow fragments to migrate toward the positive electrode.

  7. 7

    Analyze the resulting bands to determine the size of your amplified product.