Positive Sample Gel Electrophoresis
October 31st, 2018
Shepard Saabye

Objectives and Rationale: Use Gel Electrophoresis to test weather 10^0 Titer Lysate can be used to find phage, or if PCR is useless in our experiment.

Procedure:

• Retrieved PCR tubes from Freezer
• Added .40 mL 1x TAE to a flask
• Added 8 grams agarose to the flask
• Swirled and heated Agarose solution until dissolved
• Once warm, but not hot, added Ethidium Bromide to the solution
• Poured into the gel apparatus
• Set up electrophoresis machine
• Placed gel in the machine and added TAE until the gel was well submerged
• Added DNA Ladder and the 3 experimental DNAs into wells, as well as 3 positive controls
• Ran Electrophoresis for 40 minutes
• Imaged in UV imaging machine

Results: Found all tested samples to be negative, including the confirmed positive sample.

Analysis and Interpretations: Because the positive sample came out as negative, the whole base of using PCR to identify phages is shaky, even unlikely. We can run more tests to confirm, but based off two results, the 10^0 solution is not adequate for PCR.

Future Plans: I will be adopting Justin’s Phage and working to get to a high titer.