Team:BYU Provo/Notebook/Phage Purification/Springexp/Period4/Exp/6.19Spot Test

Phage Purification May - June Notebook: Experiments

6.19 Spot Test

I) Purpose

Test to see if there are viable phage from 6.17 CsCl Gradient

II) Expected Outcome

Plaques from the bands in the CsCl gradient where phage were purified.

III) Reagants Used

W3110 and BL21 freezer stocks

LB

T4 and T7 phage from CsCl gradient

.2% Agarose Top Agar

IV) Actual Procedure

Infect 500 µL of LB with 1 loop-full of bacterial freezer stock and put on shaker at 37^◦ C for 30 min.

Plate on an LB plate with 4.5 mL top agar.

Spot 5 µL of phage from CsCl gradient onto plate. Do not invert or move. Leave on desktop for one hour.

Place in 37^◦ C incubator overnight.

V) Results