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If your yeast isnt bubbling after 10 minutes, toss it and start over with fresh yeast

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doi: 10.4161/cc.8.23.10238 29 FuYLiuSYinSNiuWXiongWTanMet al

Briefly, 25-30 million cells were collected and washed twice with cold saline (0.9% NaCl), scraped into 1 ml of cold KPBS, and pelleted via centrifugation at 1,000g for 1.5min at 4C

The lack of a hook effect in degradation assays was particularly apparent in the presence of CRBN overexpression, where ( S )-dHTC1 induces maximal degradation at concentrations spanning more than 3 orders of magnitude (Extended Data Fig