Acetone extraction of bacteria pellet for HPLC (2022 iGEM)
Team Fudan iGEM
Published: 2022-10-11 DOI: 10.17504/protocols.io.eq2ly7peelx9/v1
Abstract
Pre-cold acetone is generally used for protein precipitation. Here, we collect the supernatant for membrane lipid analysis.
Steps
1.
Grow bacteria in in 150 mL 2xYT media overnight. Harvest by centrifugation at 4000 rpm for 10 minutes. Discard the supernatant.
2.
Add 4 times the volume -20°C acetone (3 mL in our case) to the extract, and vortex.
3.
Incubate sample at -20°C for 1 hour. 1h 0m 0s
4.
Centrifuge at 13500 rpm for 10 minutes at 4°C. Collect the supernatant without touching the protein pellet.
5.
Store the acetone extracted sample at -80°C for long term use, or -20°C for short term use.
