Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.2 Modeling phage plaque size

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Small Phage July - August Notebook: Experiments

7.29 Mutagen Concentration Test - Sixth Protocol

I) Purpose

To start modeling phage plaque sizes against several variables including, but not limited to phage particle size, agar concentration, and bacterial concentration.

II) Expected Outcome

A inverse relationship between phage plaque size and phage particle size/ agar concentration/bacterial concentration.

III) Reagents Used

IV) Procedure

1) E coli strain preparation (8.2-8.5)

We normally use E coli BL21 for our experiments with T7. The large phage group kindly provided the streak of E coli B.

E coli W3110 and K21 was streaked from frozen glycerol stock and agar stock respectively.

Because K21 streak from last step didn't work, liquid culture was prepared to amplify bacteria from agar stock. To three test tubes, 1mL of LB was added to each:

- test tube 1: a pipet tip was used to stab the agar stock, after this LB was pipetted up and down to get the agar into LB.
- test tube 2: a wooden stick was used to scrap the agar stock, after which it was submerged in LB
- test tube 3: after preparing test tube 2, the wooden stick used was submerged directly in the LB of test tube 3

2) Phage viability/infection test (7.29)

V) Results

VI) Conclusion