Shaker, benchtop, orbital incubator

A. Preculture preparation. A 250-mL Erlenmeyer flask containing 50 mL of mineral salt broth (MSB) (Note 1) with 0.2% L-arginine hydrochloride (L-arginine-HCI) and fitted with a cotton plug is sterilized (Note 2). In a laminar flow hood, a single colony of Pseudomonas putida 39/D (Pp 39/D) (Note 3) is selected from a fully grown plate (Notes 4, 5, 6) and transferred to the solution with a sterile loop taking care not to place the hand over the plate or flask. The flask is then placed in a benchtop orbital incubator shaker at 30°C and 200 rpm for 24 hr. 

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