Hi everyone,
I am planning to do enzyme assays for three enzymes (Isocitrate lyase, Methylisocitrate lyase, Propionyl-CoA carboxylase ) in Vibrio bacteria.
I would like to know if there is any general method for cell preparation and storing for these three enzymes?
Can I just centrifuge, wash the cells, snap free in liquid nitrogen and store in -80oC for later cell extraction and enzyme assays?
Thank you so much.
Best regards,
#Dinesh Kumar. So, after harvesting the bacteria, can I just snap freeze sample in liquid nitrogen and store at -80? Or do I need to add some chemicals for storing? Thank you so much.
Dear thao,
Your Vibrio strains should be conserved at -80°C in tryptic soy broth with 2% NaCl plus 15% (v/v) glycerol.
Best Luck.
in the link below you find , appropriate answer
https://www.thermofisher.com/sd/en/home/industrial/microbiology/microbiology-learning-center/storing-bacterial-samples-optimal-viability.html
Preparation of bacteria
Prepare a master plate of selected bacteria at least 2 days before infection. The bacteria line of 15% glycerol stock is stored indefinitely at -80 ° C. Store the master plate at 4 ° C for up to 2 weeks. Always consecutive straight main plates of frozen glycerol stock. Avoid the serial passage of bacteria from plate to plate, as repeated passage in culture can cause bacteria to develop virulent virulence.
Note: This example makes use of the Providencia morphological and Escherichia coli
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