Oct 11, 2022
Acetone extraction of bacteria pellet for HPLC (2022 iGEM)
Forked from Acetone precipitation of proteins
- igem 1
- 1Fudan University
- 2022
Protocol Citation: igem 2022. Acetone extraction of bacteria pellet for HPLC (2022 iGEM). protocols.io https://dx.doi.org/10.17504/protocols.io.eq2ly7peelx9/v1
License: This is an open access protocol 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
Protocol status: Working
We use this protocol and it's working
Created: October 10, 2022
Last Modified: October 11, 2022
Protocol Integer ID: 71123
Keywords: acetone extraction of bacteria pellet, supernatant for membrane lipid analysis, acetone extraction, membrane lipid analysis, protein precipitation, bacteria pellet, extraction, hplc, protein, bacteria
Abstract
Pre-cold acetone is generally used for protein precipitation. Here, we collect the supernatant for membrane lipid analysis.
Troubleshooting
Grow bacteria in in 150 mL 2xYT media overnight. Harvest by centrifugation at 4000 rpm for 10 minutes. Discard the supernatant.
16h
Add 4 times the volume -20°C acetone (3 mL in our case) to the extract, and vortex.
15m
Incubate sample at -20°C for 1 hour. 01:00:00
1h
Centrifuge at 13500 rpm for 10 minutes at 4°C. Collect the supernatant without touching the protein pellet.
10m
Store the acetone extracted sample at -80°C for long term use, or -20°C for short term use.