Team:BYU Provo/Notebook/Phage Purification/Winterexp/Period1/Exp/5.20Titer

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Phage Purification May - June Notebook: Experiments

5.20 Titer

I) Purpose

Test T4 and T7 phage to see if PEG purification was succesful.

II) Expected Outcome

Plaques will appear on the plates proving the presence of viable phage.

III) Reagants Used

T4 purified from

T7 purified from

2x top agar

LB

W3110 freezer stock

BL21 freezer stock

IV) Actual Procedure

Place two loop fulls of W3110 and BL21 freezer stock in separate tubes with 25 mL LB.

Place in 37° C for 1 hour.

Prepare top agar by using one part 2x top agar and one part LB to create a 1x top agar solution.

Add 4.5 mL 1x top agar to .5 mL W3110 and .5 mL BL21 and plate.

Peform a dilution series on T4 and T7 phage diluting it down to -5 concentration.

Spot dilution series T4 and T7 phage on plates.

Leave overnight in 37° C.

V) Results

Our BL21 plate with the T7 phage showed plaques. Unfortunately, these plaques were smeared and we were unable to differentiate the concentration. However, the plaques showed that our purification was successful. The W3110 plate with T4 phage showed no plaques. It did have signs of possible contamination so we will rerun the test again.