Biotechnology for Young Scientists

Cell lysis solution

Dissolve the cell lysate powder in 500mL of deionized water and stir until the solution is clear.

TBE buffer

Dilute 10mL of 10X buffer with 90mL of deionized water. This resulting 100mL solution is sufficient to make a gel

Agarozi gel

Step 1: Add 0.8 g of agarose in 100mL of 1X buffer to a 250mL beaker. Heat and stir until the solution is clear. Allow to cool to 55 degrees Celsius.

Step 2: Ready agarose gels can be stored at room temperature. Prior to electrophoresis, place in a microwave oven for 1 minute until completely liquefied.

DNA labeling solution

Dissolve 10 mL of the concentrated label solution in 90 mL of hot tap water solution.

DNA extraction from cells and labeling

Step 1: Place 10mL of dissolved onion cell solution in a test tube.

Step 2: Hold the test tube at a 45 degree angle and slowly add 5-7mL of frozen ethanol. Ethanol will form a second layer on top of the solution. Leave the solution for 2-3 minutes. You will see DNA floating in the alcohol layer.

Step 3: Bring the DNA vaccination tube to the interface of the two layers and stir.

Step 4: Carefully lift the inoculum tube and place the DNA sample on the rim of the test tube. when DNA is stretched it forms thin threads,

Gel electrophoresis

This Activity aims at assembling the electrophoresis device, forming wells and finally electrophoresis and DNA observation.

The technique of electrophoresis involves the separation and movement of DNA molecules in a gel which is exposed to an electric field. DNA is a negatively charged molecule and so the DNA fragments move towards the positive pole. Small fragments move faster than large ones.