Notebook – Protocol – 11. Cultivation

1. From plate of E. coli BL21/DE3 harboring appropriate plasmid, colony was picked and cultured in 2 ml LB medium containing antibiotic chloramphenicol (Cam) with final concentration of 34 ug/ml. 10 ml size glass tube was used. This was pre-culture.

2. The pre-culture was shaken over-night at room temperature.

3. Main culture was prepared for the next day, which contains pre-culture, LB medium, Cam, expression reagent for the expression of reporter gene mRFP (autoinduction medium which includes 20x NPS, 50x 5052 and 1,000x Mg), sugar for catabolite repression induction (either glucose, fructose, glucose+fructose, or honey). Detail receipt with various concentration of sugars was given in Tables below.

4. Main culture was shaken over-night at room temperature.

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