Endy:STN Chemostat Protocol

contact: Caitlin Conboy, Jennifer Braff

Chemostat Design
See Chemostat Design.

Day 0
Set up o/n culture in supM9 media in a small flask at 37 C.

Day 1

 * Take OD600 of o/n culture. Dilute back in the morning into 25 mL/chamber supM9 (~1:100)
 * When culture reaches ~OD 2.0 to add 20 mL to each chamber (usually late afternoon).
 * Run at 37 C. The bubbler should be set to the white dot for 6 chambers (med/high) and the effluent needle should be set to around the 24 mL mark to maintain a volume of 20 mL.
 * Add the media input (teal, 23-G1 needle) and turn on pump (currently 10 rpm for 20 mL/chamber). After about 1 hour, check that the culture volume is 20 mL and adjust effluent as necessary.
 * The cultures should wash out to a stable OD600 by the morning (day 2). OD600 should be monitored throughout the day (~every 3 hrs) and samples drawn for steady-state measurement either in the latter part of day 2 or on day 3.