Cultivation of overnight suspension culture

Cultivation of overnight suspension culture

Instruments: alcohol lamp, inoculation loop
Pharmaceutical reagents: E. coli strain (JM109)
LB broth: 1% (w/v) Bacto tryptone, 0.5% Bacto yeast extract, 1% NaCl, adjusted to pH 7.0 with 5 N NaOH, then sterilized by moist heat.
LB agar plates: LB medium supplemented with 1.5% (w/v) Bacto agar.

Method steps:
1) Each group should have a 15 mL test tube containing 2 mL of LB medium, please write JM109, and indicate the group and date.
2) Burn the inoculation ring red on the flame of the alcohol lamp, and the upper part of the ring is also burned with the flame. Allow the loop to cool and scrape off the colonies and replace the Petri dish lid.
3) Open the cap of the test tube (hold the test tube in one hand and unscrew the cap with the little finger of the hand holding the inoculation loop), and place the mouth of the tube over the alcohol lamp. Dip the colony-stained inoculation loop into the medium and tap the tube wall to dislodge the colonies. Put the cap back on after over-firing the nozzle on the alcohol lamp again.
4) The inoculation loop is sterilized by burning red on the flame and then placed back on the laboratory table.
5) Unscrew the cap of the test tube slightly, and fix the cap with tape to prevent the cap from loosening and falling off when shaking the culture.
6) Place the test tube in a 37°C incubator or water bath for overnight incubation with shaking.
7) Return the petri dish with JM109 to the teaching assistant.
Any containers or media that have come into contact with bacteria must be sterilized after use before discarding. If the bacteria is accidentally splashed during the experiment, immediately rinse with plenty of water and disinfect with 70% alcohol. When there is bacterial liquid on the table or the ground overturned, it must be cleaned with 10% bleaching water.