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

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Discussion Overview

The discussion centers around the extraction and measurement of lycopene production from E. coli using various solvents and methods. Participants are exploring practical tips to minimize solvent evaporation during absorbance measurements in 96-well microplates.

Discussion Character

  • Exploratory, Technical explanation, Debate/contested, Experimental/applied

Main Points Raised

  • One participant describes using acetone for lycopene extraction but faces issues with evaporation affecting absorbance measurements.
  • Another participant references a paper on lycopene extraction from tomato skin, suggesting it may not be directly applicable to E. coli.
  • A participant expresses the need for an organic solvent that can penetrate E. coli cell walls, is transparent, and has a higher boiling point than acetone.
  • One suggestion made is to refrigerate the samples to slow down evaporation.

Areas of Agreement / Disagreement

Participants have not reached a consensus on the best method to prevent evaporation or the most suitable solvent for lycopene extraction from E. coli. Multiple competing views and suggestions remain in the discussion.

Contextual Notes

Participants have not fully resolved the issue of solvent choice or evaporation techniques, and there are limitations in the details provided about methods used in the literature.

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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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