Mops minimal media kit

Manufactured by Teknova

The MOPS Minimal Media kit is a laboratory product designed for the preparation of a minimal growth medium. The kit contains the necessary components to create a basic medium that supports the growth of a variety of microorganisms. The kit provides a standardized and convenient way to prepare this type of media for laboratory applications.

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Lab products found in correlation

Etest strips, by bioMérieux (2 mentions) Aztreonam, by Merck Group (1 mentions) Fosfomycin, by Merck Group (1 mentions) Vancomycin, by Thermo Fisher Scientific (1 mentions) D cycloserine, by Merck Group (1 mentions) Mops ez rich, by Teknova (1 mentions)

3 protocols using mops minimal media kit

Growth Conditions for E. coli and B. subtilis

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E. coli and B. subtilis growth and media conditions were similar to those described previously^{9 (link), 20 (link)}. Briefly, E. coli were grown on LB agar with an antibiotic as needed at 37°C overnight before being inoculated into LB or 2XYT liquid medium with an antibiotic as needed and grown at 37°C.
B. subtilis cultures were inoculated from single colonies and were grown in LB Lennox and S7 liquid media as indicated at 37°C. For the preparation of S7 growth media, we modified the Teknova MOPS Minimal Media kit (M2106) to the appropriate concentrations and supplemented with 0.1% w/v glutamic acid and 40 μg/mL L-tryptophan.
For FRET experiments, single colonies grown on LB Lennox agar overnight were inoculated into S7 media and grown in a roller drum to early log phase. For imaging purposes, the cultures were then concentrated via centrifugation (3000 rcf, 2 min) and resuspended in the original supernatant.
For antibiotic treatment experiments, 3ml cultures were grown to midlog (OD₆₀₀~0.4) in S7 media before being split and treated with the antibiotic at the concentration indicated for 1.5 h. Antibiotic sensitivity experiments were performed using ETest® strips (Biomerieux) on S7 agar plates at 37°C overnight.

Zheng C.R., Singh A., Libby A., Silver P.A, & Libby E.A. (2021). Modular and single-cell sensors of bacterial Ser/Thr kinase activity. ACS synthetic biology, 10(9), 2340-2350.

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Microscopy Assay for Single-Cell Growth

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Cell cultures were grown from an individual colony in LB Miller medium (LB) to the exponential phase with an appropriate selection of antibiotics. Strains with plasmids were grown with 50 μg/ml carbenicillin during overnight growth. For microscopy, cells were then washed and diluted into LB, MOPS Rich Defined Medium (RDM; MOPS EZ Rich Defined Medium, M2105, Teknova), or defined MOPS minimal medium (MM) (MOPS Minimal Media Kit, M2106, Teknova) without antibiotic. We added 0.02–0.4% (w/v) glucose to RDM and MM, depending on the experiment (Appendix Tables S1 and S2). Cells were grown at 30°C in liquid media in a shaking incubator. To monitor single‐cell growth without division, we inhibited cell division either by inducing sulA with 1 mM IPTG from plasmid pDB192 or by adding 10 μg/ml aztreonam (Sigma‐Aldrich, A6848) shortly before microscopy (Appendix Tables S1 and S2). To inhibit cell wall synthesis, we treated cells with vancomycin (Fisher Scientific, BP2958‐1), Fosfomycin (Sigma‐Aldrich, P5396‐1G), or D‐cycloserine (Sigma‐Aldrich, 30020‐1G). To perform microscopy, we used three different supports, flow chambers, agarose pads, or donut‐shaped chambers, as described in the following. During sample preparation and subsequent imaging, the temperature was maintained at 30°C.

Oldewurtel E.R., Kitahara Y., Cordier B., Wheeler R., Özbaykal G., Brambilla E., Boneca I.G., Renner L.D, & van Teeffelen S. (2023). Cell envelope growth of Gram‐negative bacteria proceeds independently of cell wall synthesis. The EMBO Journal, 42(14), e112168.

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Comparative Growth and Antibiotic Response

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E. coli and B. subtilis growth and media conditions were similar to those described previously 10, (link)24 (link) . Briefly, E. coli were grown on LB agar with an antibiotic as needed at 37°C overnight before being inoculated into LB or 2XYT liquid medium with an antibiotic as needed and grown at 37°C. B. subtilis cultures were inoculated from single colonies and were grown in LB Lennox and S7 liquid media as indicated at 37°C. For the preparation of S7 growth media, we modified the Teknova MOPS Minimal Media kit (M2106) to the appropriate concentrations and supplemented with 0.1% w/v glutamic acid and 40 μg/mL L-tryptophan.
For FRET experiments, single colonies grown on LB Lennox agar overnight were inoculated into S7 media and grown in a roller drum to early log phase. For imaging purposes, the cultures were then concentrated via centrifugation (3000 rcf, 2 min) and resuspended in the original supernatant.
For antibiotic treatment experiments, 3ml cultures were grown to midlog (OD600~0.4) in S7 media before being split and treated with the antibiotic at the concentration indicated for 1.5 h.
Antibiotic sensitivity experiments were performed using ETest® strips (Biomerieux) on S7 agar plates at 37°C overnight.

Zheng C.R., Singh A., Libby A., Silver P.A., & Libby E.A. (2021). Protein sensors of bacterial kinase activity reveal antibiotic-dependent kinase activation in single cells.

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