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文献和实验with a strain that already has one or more plasmids use liquid drop-out media lacking the appropriate nutrient.b Incubate at 30o C in roller drum at top speed overnight. b 2. Measure the OD600 of the overnight culture.c Make an appropriate dilution
) at the final concentration of 1 tablet/50ml): PBS Buffer A: 10 mM Hepes, pH 7.9, 10mM KCl, 1.5mM MgCl2, 0.5mM DTT S1 solution : 0.25 M Sucrose, 10 mM MgCl2 S2 solution: 0.35 M Sucrose, 0.5 mM MgCl2 S3 solution: 0.88 M Sucrose, 0.5 mM MgCl2
will drop some and yeast will take longer to grow. Transformation: 1) Pre-warm 50ml 2xYAPD to 30deg in 500ml flask (at least 10min). Turn on 42deg water bath (takes 30min or more). 2) Dilute O/N culture 1:10 in water and take an OD at 600nm. Use 2xYAPD
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