Team:BYU Provo/Notebook/Phage Purification/Winterexp/Period1/Exp/6.26T7Titer

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Phage Purification May - June Notebook: Experiments

6.26 Titer

I) Purpose

Test the concentration of purified T7 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

BL21 freezer stock

LB

1.5x top agar

T7 phage from CsCl gradient

IV) Actual Procedure

Infect 500 µL of LB with 1 loop-full of BL21 freezer stock and put on shaker at 37^◦ C for 5 hours.

Plate BL21 on an LB plate with 4.5 mL top agar.

Dilute T7 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 T7 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.