Team:BYU Provo/Notebook/Phage Purification/Springexp/Period2/Dailylog

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5/13/13

- Thankfully we don’t have to use the ultracentrifuge to achieve 10,000 g. We also won’t need to order any special centrifuge tubes, so we should be able to start experimentation on Wednesday.

-We added 1 mL of w3110 and 1 mL of BL21 to 24 mL of LB each and placed them both into the 37C incubator. After an hour of incubation we infected the BL21 Ecoli with t7 phage and left to incubate until wednesday. We will infect the w3110 tomorrow with T4

5/14/13

- Today we came in and infected 24 mL of LB with 1 mL of w3310 and put in the 37C incubator overnight.

I also infected 25 mL of w3110 with 100 µL of t4 phage and left overnight in the 37C incubator overnight.

5/15/13

- We added 2.5 ml of LB to the 24 mL of t4 infected W3110 to make a final volume of 25 mL.

- We added 92.5 µL of DNAse I and 12.5 µL of RNAse A into the 25mL of t4 infected w3110 Ecoli and the t7 infected BL21.

- We then added 50 µL of chloroform to each lysate and let it incubate for 30 min at room temperature.

- We then added .73 g of solid NaCl to both lysates and let it sit at 4 C for 1 hour.

5/16/13

-Centrifuged at 8000 rpm (10,000 g) for 15 minutes. -Removed supernatant and added phage suspension buffer

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