DETERMINE OPTIMAL 3AT CONCENTRATION Sample Clauses

DETERMINE OPTIMAL 3AT CONCENTRATION. 3AT can be used to suppress background expression from the HIS reporter gene of Y190. 3AT concentration varies among different reporter strains and ranges from 0 mM (HF7c) to 15 mM (Y190). To test the optimal concentration of 3AT, one yeast colony should be re-suspended in 10 ml of TE. 100 uI of the re-suspended yeast is spread on SD-H+0mM3AT, SD-H+5mM3AT, SD-H+10mM3AT, SD-H+15mM3AT, SD-H+25mM3AT, and SD-H+40mM3AT plates. Although 15 mM 3AT is sufficient to suppress background HIS expression of Y190, higher concentrations of 3AT (30-40 mM) are routinely used in our cDNA library screening.
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