Title: Gel Electrophoresis

Date: 24 October 2018

Rationale: After the PCR procedure from the previous lab, a gel will be made and DNA loaded into the gel. An electrical current will be run through the DNA and the gel will show the DNA strands with respect to a primer mix.

Procedure:

• 35 mL TBE and 0.7g of agarose was mixed into a flask.
• The mixture was microwaved until the solution was clear
• Once the solution cooled to a temperature of ~55 degrees Celsius, 1.7 microliters of EtBr was added to the mixture.
• The mixture was poured onto an electrophoresis plate to set. Once the gel set, the dams and comb was removed and the plate was placed in the electrophoresis apparatus (the “holes” were placed towards the positive end in order to repel the negatively charged DNA)
• 5 microliters of the protein ladder was loaded into the first lane (bottom lane on picture)
• 10 microliters of PCR Primer mix 1 + DNA  (from previous lab) was loaded into the second lane (second to bottom lane in picture)
• 10 microliters of PCR Primer mix 2 + DNA was loaded into the third lane (third from bottom in picture)
• 10 microliers of Primer mix + DNA 3 was loaded into the fourth lane (fourth from bottom in picture)
• The power source was attached to the cathode and anode, and 100 V of electricity was run through the gel for ~30 minutes.

Results/Conclusions: The gel appeared to have one thick, yellow strand and one thick, blue strand. The results did not appear to be positive for any DNA Primer mix. New soil will have to be collected or a different procedure will have to be used. Attached is a picture of the gel and computer readout.