Team:BYU Provo/Notebook/SmallPhage/Springexp/Period1/Exp/5.9 T7+ Liquid Culture Phage Concentration Test

Small Phage

March-April

May-June

July-August

September-October

To determine if the volume of phage stock added effects the liquid culture concentration result

II) Expected Outcome

We expect the same results as from 5.3. We also hope to see 1 ul produce a titer to -10.

III) Reagants Used

E. Coli BL21 overnight from 5.8; T7+ from 5.6 10 ul stock; LB from 4.5

IV) Actual Procedure

1. Liquid culture set up (5.9)

Three test tubes were set up. To each, add 4mL of LB and 1mL of E coli BL21 overnight

Add 1uL, 10uL, and 100uL phage to each tube, respectively.

Liquid culture was incubated for approximately 48 hours

2. Spot test to test amplification results

Phage liquid culture was purified using procedures similar to those in 5.3 T7 phage amplification/purification. The resulted purified stock was labeled 5.13 (5.13)

Spot test was performed using procedures similar to those in 5.3 T7 phage amplification/purification (5.15)

Spot test plates were incubated overnight

V) Results

Complete clearage of liquid culture was observed for once

All spot tests showed plaques to -8, but there was also a lot of contamination -> from the LB plates; the plates have been completely disposed of.