Aug 05, 2020
Metabolite Extraction
- 1domnic colvin
- UM-DAE Centre for Excellence in Basic Sciences
Document Citation: avinash.kale 2020. Metabolite Extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.bjdtki6n
License: This is an open access document distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: August 05, 2020
Last Modified: August 05, 2020
Document Integer ID: 40083
Metabolite Extraction
The bacterial cultures were grown overnight in 200 ml LB media to sufficiently synthesize their metabolites. The bacterial cultures were centrifuged and washed to remove traces of remaining media. Cell pellets were then sonicated at amplitude 30%, pulse 10 seconds, gap 10 seconds, for a total time of 2 minutes: 30 seconds or 15 Cycles in 300 µL of 1x PBS. The samples were centrifuged at 12000 rpm for 20 minutes and the supernatant was separated. The metabolites were precipitated using acetonitrile in twice the volume of the supernatant by incubating at -80 °C overnight. The clear supernatant was centrifuged, transferred into new tubes, followed by air drying(27). These dried samples were further given for LCMS.