Objectives: Have a large amount of pure phage DNA pelleted, to extract and analyze.

Rationale: To analyze the genome, the phage DNA must be extracted in large quantities.

Procedure:
Note – These procedures were not done aseptically

• Added 10 mL of high titer lysate to a 50 mL conical tube
• Added 40 uL nuclease using micropipettor
• Transferred 4 mL Polyethylene Glycol to Lysate
• Left 50 mL tube in an incubator at 37 degrees Celsius for 30 minutes
• Removed tubes from incubator, and left at room temperature for 45 minutes
• Centrifuged the lysate mixture at 10,000 Gs for 20 minutes
• Removed the Supernatant
• Air dried pellets and placed in a microcentrifuge tube

Future Plans: Use DNA pellets to complete extraction and PCR procedures.