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- Overview
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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 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
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