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Preprint

100x Trace Elements for M9 minimal medium

AF Avi Flamholz
ED Eli Dugan
RM Ron Milo
DS David F Savage

Last updated date: Jul 13, 2021 Views: 3988 Forks: 0

original An abbreviated version of this protocol was published in eLife in Oct, 2020
Functional reconstitution of a bacterial CO2 concentrating mechanism in Escherichia coli
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Intro

Recipe is for 100x Trace Elements solution used for liquid and solid media. 

Materials

  • EDTA powder
  • NaOH 1 M stock (for pH of the EDTA solution)
  • FeCl3-6H20 powder 
  • ZnCl2 
  • 0.1 M CuSO4 stock 
  • 0.2 M CoCl2 stock 
  • 0.1 M H3BO3 stock 
  • 1 M MnCl2 stock 

 

Concentrations 

Final concentrations are:

ComponentMass ConcentrationMolar Concentration (uM)Final Concentration in Media (uM)
EDTA5 g/L1.3e+41.3e+2
FeCl3-6H2O0.83 g/L3.1e+33.1e+1
ZnCl284 mg/L6.20e+26.20e+0
CuSO4 - 5 H2O19 mg/L760.76
CoCl2 - 6H2O10 mg/L420.42
H3BO310 mg/L1621.62
MnCl2 - 4H2O1.6 mg/L8.10.081

 

Procedure: 

  1. Dissolve 5 g of EDTA in 800 mL milliQ.
  2. Stir and pH to 7.5 with NaOH.
  3. Keep stirring, add 830 mg of FeCl3-6H2O
  4. Add 84 mg ZnCl2
  5. Add the liquid stocks in the following volumes (table below)
  6. Add MilliQ water up to 1 L.
  7. Sterile filter in 1 L filter system.
stockstock concentrationvolume
CuSO4-5H2O0.1 M765 uL
CoCl2 - 6H2O0.2 M210 uL
H3BO30.1 M1.6 mL
MnCl21 M8.1 uL
Copyright: Copyright Flamholz et al. This article is distributed under the terms of the Creative Commons Attribution License (CC BY 4.0).
How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Flamholz, A, Dugan, E, Milo, R and Savage, D(2021). 100x Trace Elements for M9 minimal medium. Bio-protocol Preprint. bio-protocol.org/prep1287.
  2. Flamholz, A. I., Dugan, E., Blikstad, C., Gleizer, S., Ben-Nissan, R., Amram, S., Antonovsky, N., Ravishankar, S., Noor, E., Bar-Even, A., Milo, R. and Savage, D. F.(2020). Functional reconstitution of a bacterial CO2 concentrating mechanism in Escherichia coli. eLife. DOI: 10.7554/eLife.59882

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