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Saturated overnight, LBH cultures were diluted 100-fold in LBH medium and grown at 37°C for 1.5 hours, at which time doxycycline (100 ng/ml) was added to induce protein production. After 1 hour of induction, cells were plated on LBH solid medium containing doxycycline (10 ng/ml), and the plated cells were irradiated with different doses of UVC light using a Stratalinker 1800 UV crosslinker (Stratagene) and then incubated in the dark overnight at 37°C for colony quantification. Control cultures without doxycycline induction were treated identically.
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