Rationale:

Running a gel electrophoresis this class to test Soil Sample C. The gel will show if there are phages in the soil sample with bands. If bands show up on the lane in line with the location where the enriched lysate was pipetted into, the soil sample contains phages. If nothing shows up then that represents the absence of phage.  

Procedure:  

1. Using 70% Ethanol and Cidecon created an aseptic zone.  
2. A Erlenmeyer flask was obtained and 40 mL of TBE was added.  
3. Using a weight boat 0.8 grams of Agarose was measured.  
4. Heated the solution in the microwave, and mixed until the solution turned clear.  
5. Waited until the solution was cooled, and then pipetted 2 uL of Ethidium bromide to the solution  
6. Mixed the solution and poured into the gel trays.  
7. Inserted the comb at one end and waited for the gel to set.  
8. After 20 minutes, the gel had set, and was transferred to the electrophoresis box.  
9. 1X TBE was added to fill the electrophoresis box until TBE covered the gel.  
10. The small PCR tubes were obtained from the fridge and 2.5 uL of dye was added to each of the primers and positive controls.  
11. 10 uL of solution in PCR tubes were added to respective place in gel.  
12. 5 uL of DNA ladder was added into the middle of the gel.  
13. Primer 1, Primer 2, Primer 3, Ladder DNA, Positive Control 1, Positive Control 2, and Positive Control 3 were pipetted into the gel in this order.  
14. The gel electrophoresis box was attached to the power source and let to run for 45 minutes on 100 V.  
15. After 45 minutes, gel with the tray was removed and scanned.  
16. Results were viewed and recorded.  

Observations/ Results: 

The results of the gel electrophoresis was negative. This tells that Soil Sample C did not have phage.

Next Steps/ Conclusion: 

This process might not be providing the desired results as previously tested soil samples that were positive resulted in negative gels as well. Since our gel came out negative, we are going to try to create a high titer plate in our group using the plaque and enriched lysate of the person who found a plaque previously.