Team:BYU Provo/Notebook/LargePhage/Springexp/Period1/Exp/Team:BYU Provo/Phage Liquid Culture Mutagenesis
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'''I) Purpose''' | '''I) Purpose''' | ||
- | : | + | : The purpose of this experiment is to grow T4 phage in the presence of 5’-bromodeoxyuridine, inducing mutations as the phage replicates. |
+ | : | ||
'''II) Expected Outcome''' | '''II) Expected Outcome''' | ||
- | : | + | : This mutagenesis should produce some phage with multiple genomes packed inside of elongated capsids. |
+ | : | ||
- | '''III) | + | '''III) Materials''' |
- | : | + | : E. coli bacteria at density of 3.0 x 10^8 cfu/mL (OD600 of __) |
+ | : Adenine | ||
+ | : LB Broth (rich media) | ||
+ | : 5-bromodeoxyuridine | ||
+ | : M9+ media (minimal media) | ||
+ | : Chloroform | ||
+ | : High titer phage lysate of T4 stock | ||
- | '''IV) | + | '''IV) Methods''' |
- | + | : 1. In a flask, start a culture of E. coli in rich media (LB) supplemented with 20 ug adenine per mL. Grow to a density of 3.0 x 10^8 cells/mL. (OD600 reading of ___) | |
- | : | + | : 2. Chill and pellet the E. coli. Then resuspend at a concentration of 1.5 x 10^8 cells/mL in M9+ medium supplemented with the following (per mL): 200ug 5-bromodeoxyuridine, 20 ug uracil, 20 ug adenine. |
- | : | + | : 3. To a 30 mL amount of resuspended E. coli, add 2 x 10^9 phage particles and aerate until cleared. |
- | : | + | : 4. Chill and pellet the bacteria, transfer to a new tube, and add chloroform to kill any remaining E. coli. Store at +4C. |
'''V) Results''' | '''V) Results''' |
Latest revision as of 21:49, 5 June 2013
Large Phage May - June Notebook: Experiments
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Title
I) Purpose
II) Expected Outcome
III) Materials
V) Results
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