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

Phage Purification

March-April

May-June

July-August

September-October

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