Preparation of seed culture fluids and preparation of fermentation tanks

materials, equipment, andreagents

1. Ultra-net work

, Reagents

"

I,"> media preparation

2, fermentation medium

3, 100 mmol/L IPTG storage liquid

4, 10% phosphoric acid

5, 40% sodium hydroxide solution: 121 degrees C autocultension 20min.

6, bubble enemy: 121 degrees C autocultension 20min standby

2, seed culture preparation

frozen glycera bacteria, according to 2.5% to 3% of the inoculation volume into a bottle containing 100 ml LB medium, 37 degrees C, 200rpm/min overnight culture into the fermentation tank, inoculation concentration of 200ml/3L.

3, sterilization of fermentation tanks

1. Pour the preparation of the fermentation medium into the fermentation tank.

2． Turn on the power switch on the fermentation tank system, select "fermentation" and adjust "screen" to "calibration" to confirm with "enter".

3． Corrects the zero and full scale of the pH electrode.

1) with" to "zero".

4． Corrects the dissolved oxygen electrode zero.

5． Each electrode is inserted into the corresponding hole in the top cover plate, tighten the nut, insert the pH electrode with great care, if necessary, add some distilled water lubrication, to prevent electrode damage.

6． Use a rubber band to hold each air import and exit of the control reagent bottle.

7． Remove the connecting cables on each electrode and put on your own protective sleeve. Unplug the airtrade at the entrance to the filter and seal the seal film. Pull out the in and out pipes of the air-out condenser.

8． Remove the cooling pipe and remove the mixing motor. Remove the fermentation tank from the base.

9． Fermentation tank (internal medium), reagent triangulation bottle in a high-pressure sterilization pot sterilization, 121 degrees C autocultic 30min.

10． After sterilization is over, after cooling, the fermentation tank is relocated to the base for standby.