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

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Revision as of 20:11, 5 August 2013

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

LB
Different stock of E coli strain.

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 pipette 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 (8.5)

- Prepare 2 plates for a spot test using E. coli BL21 by adding 0.5mL overnight into a tube, mixing it with 5mL of x1 top agar, and then plating it. Repeat for E. coli B and E. coli W3110.

-Spot 5ul each of T1, T3 30, T3 14, T5, and T7 onto one plate of each bacteria. Spot T2 10, T2 50, T4, x174 20, and x174 40 onto the other plate of each bacteria.

V) Results

VI) Conclusion

@@ Line 59: / Line 59: @@
 * 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 1: a pipette 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
-) Phage viability/infection test (7.29)
+) Phage viability/infection test (8.5)
+: - Prepare 2 plates for a spot test using E. coli BL21 by adding 0.5mL overnight into a tube, mixing it with 5mL of x1 top agar, and then plating it. Repeat for E. coli B and E. coli W3110.
+: -Spot 5ul each of T1, T3 30, T3 14, T5, and T7 onto one plate of each bacteria. Spot T2 10, T2 50, T4, x174 20, and x174 40 onto the other plate of each bacteria.
 '''V) Results'''