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Team:BYU Provo/Notebook/SmallPhage/Springexp
From 2013.igem.org
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May 1 - May 12
This is the start of Spring Term at BYU! The weather is finally getting better: no more snowing. And we are ready to do some serious lab work! While we are waiting for our mutgen, 5-bromodeoxyuridine, to arrive, we decided to start this term with establishing the basic procedures for our direct evolution method. Specifically, this involves making sure that we can (1) grow and amplify T7 phage in liquid culture, (2) select for smaller phage using concentrated top agar, and (3) amplify phage from a single plaque. During these two weeks, we also designed primers for cloning and sequencing T7 capsid proteins as well as mapped out detailed procedures for both cloning T7 capsid proteins and applying mutagen.
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| May 13 - May 26
In these two weeks, we performed our first round of mutagenesis and first PCR. Although we are still testing the results to optimize our protocol, we are optimistic about the outcome!
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| May 27 - June 16
We discovered that our first round of mutagenesis didn't work properly so we adjusted it slightly and performed a second round. We also sequenced the T7 capsid protein.
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| June 17 - June 30
We completed our second round of mutagenesis, but to no avail. There was no evidence of mutation so we started a third round of mutagenesis by altering our protocol a little bit.
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