Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.16 Modeling phage plaque size

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Small Phage July - August Notebook: Experiments



Overview
March-April
May-June
July-August
September-October

8.16 Modeling phage plaque size - Experiment One


I) Purpose

II) Expected Outcome

  • A inverse relationship between phage plaque size and phage particle size/genome size.

III) Reagents Used

  • Accurate x2 top agar made using a volumetric flask.
  • LB
  • E coli B.
  • Stocks of phage: T1, T2, T3, T4, and T7.

IV) Procedure

1) Plating (8.16)

  • To see a statistical significance in plaque sizes between different phages, many plaques are needed. Thus 10 plates were used for T1, T2, T3, and T7. Because T4 form much smaller plaques, only 3 plates were used.
  • Experimental procedures are as follow: 0.5mL of E coli BL21 (OD 0.5, adjusted after initial measurement of 0.558) was infected with 20uL of respective phage dilution samples for 10 minutes. 5mL x1 top agar was then added to each test tube, and the content was plated. The plates were incubated for 24 hours.

V) Results

1) Plating

# Results
Phage Dilution used Average plaques/plate Average diameter Total plaques analyzed Reported capsid size Reported genome size
T1 -2 18 w/ overlap -3 0
T2 -4 22 w/ overlap -5 6
T3 -5 33 w/ overlap -6 7
T4 -6 a lot w/ overlap -7 47
T7 -4 a lot w/ overlap -5 18


VI) Conclusion