Team:BYU Provo/Notebook/Phage Purification/Winterexp/Period1/Exp/5.22T4Titer

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Phage Purification May - June Notebook: Experiments

5.22 T4 Titer

I) Purpose

Retest T4 phage to see if PEG purification was succesful.

II) Expected Outcome

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

III) Reagants Used

T4 purified from

2x top agar

LB

W3110 freezer stock

IV) Actual Procedure

Place two loop fulls of W3110 freezer stock in 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 plate.

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

Spot dilution series T4 phage on plate.

Leave overnight in 37° C.

V) Results

Our titer once again showed contamination on the plate. However, we were able to see plaques forming on the plate thus confirming the presence of viable T4 phage.