Lab 4: Gel Electrophoresis
To determine the presence or absence of PCR products and quantify the size (length of the DNA molecule) of the product.
Students will use agarose gel electrophoresis to determine the presence and size of both the arthropod COI barcoding gene and the Wolbachia 16S rRNA gene amplified by PCR. Upon completion of this activity, students will have integrated scientific discovery, inquiry and biotechnology. Students will understand how PCR products are visualized and analyzed based on size.
Teaching Time: One class period if using MiniOne or MiniPCR; Two class periods if using standard gel electrophoresis
Recommended Project Guide
The MiniOne System utilizes a built-in blue light and photo hood to allow real-time visualization of the bands as they move down the gel. Gels can be poured, run, and visualized in a single class period. For best results, we recommend using the MiniOne GreenGel GelCups (1% TBE).
Another class favorite is the MiniPCR System. It uses a similar technology to the above protocol; refer to the manufacturer guide to adjust for slight differences with the equipment.
Standard Gel Electrophoresis
This protocol can be used for most standard gel rigs. It will take generally take one class period to prepare and cast the gel, and a second class period to run and visualize the gel. Note that GelRed (or GelGreen) is added to the gel before it is cast. If you prefer post-staining the gel, you will need to allow an extra 30 min after the gel run to stain and image the gel. Refer to manufacturer guide for specific instructions. Most important: make sure the DNA Stain is compatible with your transillumator (blue light or UV).