Team:BYU Provo/Notebook/Phage Purification/Summerexp/Period1/Exp/7.26 Phage Titer

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Phage Purification July - August Notebook: Experiments

Phage Purification

7.26 Titer

I) Purpose

Test the concentration of purified T4 phage.

II) Expected Outcome

Plaques from the bands in the CsCl gradient where phage were purified. Expecting plaques down to at least -5 concentration

III) Reagants Used

E Coli B freezer stock

LB

1.5x top agar

T4 phage from CsCl gradient

IV) Actual Procedure

Infect 500 µL of LB with 1 loop-full of E. Coli B freezer stock and put on shaker at 37^◦ C for 5 hours.

Plate w3110 on an LB plate with 4.5 mL top agar.

Dilute T4 using a dilution series.

Spot 2 µL of phage from dilution series onto plate using the concentrations 0, -2, -3, -4, -5, and -6.

Place in 37^◦ C incubator overnight.

V) Results

The titer of T4 phage showed plaques down to a -5 concentration. This concentration will allow us to perform and EM and examine how pure the phage solution is.

T4 Titer