Team:BYU Provo/Notebook/SmallPhage/Springexp/Period1/Exp/Team:BYU Provo/5.6 T7+ Liquid Culture Phage Concentration Test


Small Phage May - June Notebook: Experiments

Small Phage

5.6 T7+ Liquid Culture Phage Concentration Test

I) Purpose

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

II) Expected Outcome

We hope that 100uL of phage will be enough to increase the liquid culture concentration

III) Reagants Used

E. Coli BL21 overnight from 5.5; T7+ from 5.3 T7 phage amplification/purification; LB from 4.5

IV) Actual Procedure

Liquid culture set up (5.6)

- Three test tubes were set up. To each, add 4mL of LB and 1mL of E coli BL21 overnight
- Add 10uL, 100uL, and 1mL to each tube, respectively.
- Liquid culture was incubated for approximately 48 hours (5.6 4:00pm - 5.8 6:00pm)

Spot test to test amplification results (5.8)

- Phage liquid culture was purified and spot test was performed using procedures similar to those in 5.3 T7 phage amplification/purification
- The purified stock was labeled 5.6
- Spot test plates were incubated overnight

V) Results (5.9)

- The 10 ul spot test went down to -9, 100 ul to -8, and 1 ml to -7
- Thus, 10 ul yields the highest titer
5.10 phage concentration test.JPG