https://wiki.yeastgenome.org/index.php?action=history&feed=atom&title=UW-Stout%2FUltraviolet_Light_FA22 2026-09-06T02:25:41Z Revision history for this page on the wiki MediaWiki 1.31.14 https://wiki.yeastgenome.org/index.php?title=UW-Stout/Ultraviolet_Light_FA22&diff=405540&oldid=prev 2022-12-15T20:32:43Z

<p>Created page with &quot;==Methods (Calibration Experiment)== ===Materials=== *60mm plates with YPD media *Yeast to be tested (suspended in media) *Phosphate buffered saline (1X, pH 7.4) *Glass beads...&quot;</p> <p><b>New page</b></p><div>==Methods (Calibration Experiment)==<br /> ===Materials===<br /> *60mm plates with YPD media<br /> *Yeast to be tested (suspended in media)<br /> *Phosphate buffered saline (1X, pH 7.4)<br /> *Glass beads for plating<br /> <br /> ===Equipment===<br /> *Spectrophotometer<br /> *Model LS-100-3 UV Exposure System<br /> <br /> <br /> ===Procedure===<br /> ====Sample Prep====<br /> #Use a spectrophotometer to measure the optical density of the stock yeast at 600 nm.<br /> #Dilute a sample to 25 µL at a concentration of 1 cell/µL using phosphate-buffered saline. Vortex to mix.<br /> #Pour 4-8 beads on the plate.<br /> #Pipette 25 µL of dilute cells onto the plate.<br /> #Shake to spread the sample on a petri dish.<br /> #Dispose of glass beads.<br /> <br /> ====UV Exposure====<br /> #Input sample plates into the UV exposure system at 400 watts for 10, 20, 50, 100, 200, 300, 400, 500, and 600 seconds.<br /> #Incubate plates at 30 °C in the dark for 24-48 hours.<br /> #Measure the number of colonies.<br /> <br /> <br /> ==Calibration Experiment Data==<br /> '''The number of colonies on each plate:'''<br /> *10 seconds - 18 colonies<br /> *20 seconds - 19 colonies<br /> *50 seconds - 12 colonies<br /> *100 seconds - 11 colonies<br /> *200 seconds - 12 colonies<br /> *300 seconds - 12 colonies<br /> *400 seconds - 6 colonies<br /> *500 seconds - 1 colony<br /> *600 seconds - 0 colonies<br /> <br /> <br /> <br /> ===Calibration Experiment Interpretation===<br /> Based on the data provided by the calibration experiment, the knock-out experiment protocol will utilize a 400-second exposure period. This period was shown to be a middle point in the number of colonies killed off. <br /> <br /> <br /> ==Knock-out experiment Protocol==<br /> ===Materials===<br /> *60mm plates with YPD media<br /> *Yeast to be tested (suspended in media)<br /> *Phosphate buffered saline (1X, pH 7.4)<br /> *Glass beads for plating<br /> <br /> ===Equipment===<br /> *Spectrophotometer<br /> *UV exposure system<br /> <br /> <br /> ===Procedure===<br /> ====Sample Prep====<br /> #Use a spectrophotometer to measure the optical density of the stock yeast at 600 nm.<br /> #Dilute a sample to 25 µL at a concentration of 4 cells/µL using phosphate-buffered saline. Vortex to mix.<br /> #Pour 4-8 beads on the plate.<br /> #Pipette 25 µL of dilute cells onto the plate.<br /> #Shake to spread the sample on a petri dish.<br /> #Dispose of glass beads.<br /> <br /> ====UV Exposure====<br /> #Input sample plates into the UV exposure system at 400 watts for 400 seconds.<br /> #Incubate plates at 30 °C in the dark for 24-48 hours.<br /> #Measure the number of colonies.</div>

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