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Dorman:P22 transduction: Difference between revisions
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'''Day 1''' | '''Day 1''' | ||
# Set up multiple transductions: | # Set up multiple transductions: | ||
* 100 μL of P22 lysate + 100μL of recipient strain | #* 100 μL of P22 lysate + 100μL of recipient strain | ||
* 100 μL of 1:10 diluted P22 lysate + 100μL of recipient strain | #* 100 μL of 1:10 diluted P22 lysate + 100μL of recipient strain | ||
* 100 μL of 1:100 diluted P22 lysate + 100μL of recipient strain | #* 100 μL of 1:100 diluted P22 lysate + 100μL of recipient strain | ||
* 100 μL of recipient strain (no phage control- confirms that there are no mutants in the recipient population) | #* 100 μL of recipient strain (no phage control- confirms that there are no mutants in the recipient population) | ||
* 100 μL of P22 lysate (no recipent control- confirms that the lysate is not contaminated with bacteria.) | #* 100 μL of P22 lysate (no recipent control- confirms that the lysate is not contaminated with bacteria.) | ||
#Incubate these mixtures at 37°C (no shaking necessary) for 1 hour. | #Incubate these mixtures at 37°C (no shaking necessary) for 1 hour. | ||
#Plate the entire volume of the transduction mixtures onto L agar plates with appropriate antibiotics. | #Plate the entire volume of the transduction mixtures onto L agar plates with appropriate antibiotics. | ||
