Team:BYU Provo/Notebook/LargePhage/Summerexp/Period11/Dailylog

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Today we tried to finish off a mutagenesis procedure.  We found that our new T4 WT Keltzie phage stock produced plaques down to between 10^-6 and 10^-9.  I used 1 mL of this lysate to start the mutagenesis procedure, adding it to 25 mL of bacteria that had an optical density of 0.155.  However, after 4 hours the bacterial cultures looked more like they had been cleared, rather than getting more turbid due to a superinfection.  Adding chloroform confirmed this--if anything, the 5 mL sample we removed and added chloroform to got more turbid rather than clearing.  This is the first time we've seen actual clearage in a liquid culture.  All of the other times we've created superinfection conditions.
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We added 0.5 mL of overnight bacterial culture to each flask to see if we can restart bacterial growth in the flask in superinfection conditions.  We'll let this incubate overnight and harvest it tomorrow.
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Revision as of 21:37, 16 October 2013


Large Phage September - October Notebook: October 16 - October 31 Daily Log



Large Phage
March-April
May-June
July-August
September-October

9/16/13

Today we tried to finish off a mutagenesis procedure. We found that our new T4 WT Keltzie phage stock produced plaques down to between 10^-6 and 10^-9. I used 1 mL of this lysate to start the mutagenesis procedure, adding it to 25 mL of bacteria that had an optical density of 0.155. However, after 4 hours the bacterial cultures looked more like they had been cleared, rather than getting more turbid due to a superinfection. Adding chloroform confirmed this--if anything, the 5 mL sample we removed and added chloroform to got more turbid rather than clearing. This is the first time we've seen actual clearage in a liquid culture. All of the other times we've created superinfection conditions.

We added 0.5 mL of overnight bacterial culture to each flask to see if we can restart bacterial growth in the flask in superinfection conditions. We'll let this incubate overnight and harvest it tomorrow.


9/18/13

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9/23/13

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9/25/13

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