Previous Results:

• The spot test (11/14) with dilutions through 10^-8 of Lysate #8 was examined. All dilutions were lysed, therefore the titer could not be calculated. Dilutions of 10^-8 will be have to be made since the titer is a high titer and can not be calculated with a spot test.

Objective:

• Complete the first step of DNA extraction by obtaining a phage precipitate

Procedure:

1. Aseptic Zone was created with CiDecon, 70% Ethanol, and an Ethanol burner
2. Lysate #8 was obtained and 10 mL was put into a 50 mL tube
3. 40 microliters of nuclease was added to the lysate and tube was inverted 10 times to mix
4. 4 mL of phage precipitate was added to 50 mL
5. Mixture was placed in 37 degrees Celsius for 30 min, then room temp for 40 min
6. The 50 mL tube was then massed (29.57 g) and a 50 mL tube was filled with water of equal mass (29.63 g) to prepare for centrifuge
7. The tube was then centrifuged at 10,000 g for 20 min, in 4 degrees Celsius
8. Precipitate was then placed in -20 degrees Celsius freezer until next lab

Results:

• Experiment was not complete, therefore there are no results to report

Next Lab:

• During next lab, the second part of DNA extraction will be completed. Further dilutions of Lysate #8 will be made to calculate the titer of the lysate and move on to a TEM.