Rationale: To prepare the gel and to create a gel electrophoresis to determine whether or not there are phages present within the sample.

Procedure:

• 0.8 grams of agarose powder was collected and poured into an Erlenmeyer flask along with 40 mL of 1X TBE.
• The flask was swirled around to make sure that the components are somewhat mixed together.
• The solution was heated until it becomes a homogenous mixture.
• The flask was left alone to cool to a warmer state.
• 2 µL of ethidium bromide was added to the warm solution.
• The agarose was poured onto the gel apparatus then the comb was inserted on the side.
• The gel was set aside to solidify for 30 minutes.
• Once the plate has solidified, the comb was gently removed and enough TBE was poured to cover the top layer of the gel within the apparatus.
• Then, 10 µL of phage buffer was inserted in the first slot then another 10 µL of phage buffer was added in another slot.
• In the next three slots, the three samples were added in increments of 10 µL.
• After the samples, 0.5 µL of DNA ladder was added.
• In the last two windows, 10 µL of phage buffer was added.
• The lid was laid on top and the power cables attached the apparatus to the power source
• The power source was put to 100 V and 4.5 mA.
• The power source ran for 30-40  minutes to ensure that the DNA had fully expressed itself.
• After the necessary time connected to the power source, the gel was taken out of the apparatus and scanned using the BioRad Gel Doc EZ Imager.

Results and Analysis:

From the BioRad Gel Doc EZ Imager, the results came back to negative. There were some positive results but that was from the phage buffers forming dimers.

Conclusion:

In order to conduct a gel electrophoresis, the gel must first be created. The agarose powder and TBE were combined in a flask and swirled. Then, the solution was heated to reach a homogenous mixture which was then left alone to cool down to a warmer condition. 2 µL of ethidium bromide was added to the solutions then quickly poured onto the gel apparatus with a comb placed on the side. The apparatus was set aside to solidify for 30 minutes. After solidifying, the comb was removed and TBE was poured to cover the surface of the gel. Four sets of phage buffers were placed, two sets on the outside. The next three sets included samples and the fourth was the control. After the samples, the ladder was filled into the slot. The lid was placed on the power source and was turned to 100 V. The power source remained on. After 40 minutes, the gels were examined.

Future Plans:

Due to the negative results from the imaging, collect more soil on 10/24/18 and wash and enrich.