Microbial Cell Procedure
Review Tissue handling precautions before proceeding.
This is a general procedure and the results can vary according to cell type. We suggest run a small trial
to determine how much culture media volume (1, 5, 10, 20 & 50 mL) is necessary to get enough biomass to generate
a suitable instrument response. PLEASE FOLLOW THE RULES FOR LABELLING.
⦁ Measure the optical density (OD600) for each culture in solution (required for LC-MS normalization).
⦁ Inactivate the cells metabolism by cooling down the samples on ice. Please note that alternative
quenching strategies can be used.
⦁ Once cooled down, centrifuge samples at 4800 g for 10 min at 4°C to pelletize the cellular mass.
⦁ If desirable, save the medium for analysis (see Cell culture medium procedure)
⦁ Wash the cell pellet with 1X phosphate buffered saline (PBS) with enough volume to cover the pellet.
⦁ Discard the PBS and centrifuge samples at 4800 g for 2 min at 4°C and remove residual medium by pipetting.
⦁ Proceed to the extraction step or snap freeze in LN2 and store at −80° C
until analyzed. If needed, all transportation should be carried out in dry ice.