Need Tips and Tricks on plate-reading Lycopene production of E.coli

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SUMMARY

The discussion focuses on optimizing the extraction of Lycopene from E. coli using acetone while addressing the issue of solvent evaporation during absorbance measurement at 470 nm in 96-well microplates. The user seeks advice on alternative solvents with higher boiling points than acetone (56°C) that can effectively penetrate E. coli cell walls. Additionally, the user inquires about methods to minimize acetone evaporation during readings, such as refrigeration of samples. The conversation highlights the need for detailed methodologies in literature regarding absorbance measurement versus HPLC evaluation.

PREREQUISITES
  • Understanding of Lycopene extraction techniques from bacterial cells
  • Familiarity with 96-well microplate absorbance measurement
  • Knowledge of organic solvents and their properties
  • Basic principles of centrifugation in laboratory settings
NEXT STEPS
  • Research organic solvents with higher boiling points suitable for E. coli extraction
  • Learn techniques to minimize solvent evaporation in microplate assays
  • Explore HPLC methods for Lycopene quantification
  • Investigate refrigeration effects on sample stability during absorbance readings
USEFUL FOR

This discussion is beneficial for undergraduate students, microbiologists, and researchers involved in microbial metabolite extraction and analysis, particularly those focusing on Lycopene production from E. coli.

koitk
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TL;DR
I need to measure the Lycopene production of E.coli strain on different mediums.
I have used Acetone, to extract Lycopene from bacteria, as described in literature. But when I started to measure the absorbance (470 nm) using 96-well microplates, some of the acetone would evaporate and ruin my results (even with the lid on).
Most of the literature that i have read uses HPLC to evaluate production, but some have used absorbance as well, but they don't specify their methods in great detail.
Can anyone help me by give me some tips, how could i slow down the evaporation. Or maybe i should use a different solvent?

Thank you in advance,
a desperate undergrad :)
 
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Thank you for your reply!
This is a very detailed description of lycopene extraction from tomatoes. But it will not be of much use to me.
Extraction of lycopene form bacterium is much easier, and i have read articles about it. I just need to add solvent, incubate and centrifuge pellet.
My main concern is finding a organic solvent, that can penetrate trough E.coli cell walls, is transparent and has a higher boiling point than acetone (56 C).
Or are there any tips, how to use acetone as a solvent and read 96-well microplates in a plate reader, without it evaporating.
 
Refrigerate the samples?
 

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