Rationale: We are preforming a PCR using Soil Sample C to test for gel electrophoresis which will test whether the soil sample shows DNA indicating the presence of phage.  

Procedure:  

1. An aseptic zone was created using Cidecon and 70% Ethanol.  
2. Enriched lysate was obtained from the fridge and poured into a 15 mL tube to be centrifuged.  
3. The enriched lysate was massed before placing in the centrifuge. 
4. Then the enriched lysate was placed in the centrifuge with a water tube with equal mass. 
5. Enriched lysate was obtained from the centrifuge and 1 mL was added to a small tube using a bulb pipette.  
6. The tube was boiled for 10 – 15 minutes.  
7. Obtained 3 PCR tubes containing 12.5 uL Taq polymerase.  
8. Added 2uL of 3 different enriched lysates.  
9. Obtained 3 different primers : primer 1 , primer 2, and primer 3 and added 4 uL of the each primer into each of the respective tubes.  
10. Obtained ddH2O water and added 2.5 uL into each of each tube.  
11. Placed the PCR tubes into the fridge to be used for gel electrophoresis. 

Observations/ Results: 

The Taq Polymerase used this time was clear, and therefore next class we will have to add the dye. A different Taq Polymerase was also used so it could produce different results for the gel electrophoresis.  

Next Steps/ Conclusion: 

Next class we will run a gel electrophoresis in order to determine if phage exists in Soil Sample C. The gel electrophoresis will display DNA lines if there is any phage present in the soil. If nothing is present then that represents the absence of phage, and a new soil sample can be tested.