K 12 mg1655

Manufactured by American Type Culture Collection

Sourced in United States

K-12 MG1655 is a laboratory strain of Escherichia coli, a common bacterium. It is a widely used model organism in microbiology and molecular biology research.

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Lab products found in correlation

Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions) Neb 5 alpha, by New England Biolabs (1 mentions) Syto 9 green fluorescent nucleic acid stain, by Thermo Fisher Scientific (1 mentions) Sybr green master mix, by Thermo Fisher Scientific (1 mentions) Tri reagent, by Merck Group (1 mentions) Caffeine, by Merck Group (1 mentions)

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MG1655 (8 citations) E. coli K-12 MG1655 cells (2 citations)

4 protocols using k 12 mg1655

Escherichia coli Strains and Phage Protocols

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The strain Escherichia coli NEB 5-alpha (New England Biolabs, NEB C2987H) was used as a cloning strain. E. coli K-12 MG1655 (ATCC 47076) was used in all experiments involving phage or toxicity. Unless mentioned otherwise, cells were grown in MMB (LB + 0.1 mM MnCl₂ + 5 mM MgCl₂, with or without 1.5% agar) with the appropriate antibiotics: ampicillin (100 μg/ml) or chloramphenicol (30 μg/ml). Expression was repressed using 1% glucose and induced using 0.02%−0.2% arabinose or 0.1–1 mM IPTG.
Phage infections were performed in MMB media with or without 0.5% agar. E. coli phages P1, T4, T5, T7 and Lambda were provided by U. Qimron. Phages SECphi17, SECphi18, SECphi27, SECphi4, and SECphi6 were isolated in the Sorek laboratory^{47 ,48 (link)}. Phages T2 and T6 were obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) (DSM 16352 and DSM 4622, respectively).

Wein T., Johnson A.G., Millman A., Lange K., Yirmiya E., Hadary R., Garb J., Steinruecke F., Hill A.B., Kranzusch P.J, & Sorek R. (2023). CARD-like domains mediate anti-phage defense in bacterial gasdermin systems. bioRxiv.

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Caffeine's Impact on E. coli Virulence

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Caffeine was purchased from Sigma-Aldrich (cat # C0750, Saint Louis, MO, United States). The bacterial strains used were CFT073, UPEC strain (ATCC # 700928; United States) and K12 MG1655 (ATCC # 700926; United States) which were first streaked onto fresh plates from glycerol stocks stored in −80 °C. Single colony was then picked and grown in Luria–Bertani (LB) (SRL Cat# 29817, India) media at 37 °C for 24 h with continuous shaking at 150 rpm to obtain primary culture. For all the experiments 10⁷ CFU/ml E.coli suspensions were used to ensure similar number of bacteria in each experiment. Other chemicals used were 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide-MTT(Invitrogen Cat # sM6494; United States), Trypsin–EDTA solution (1×) (Himedia Cat # TCL048)for cell viability of bacteria. For RT PCR, TRI Reagent (Sigma Cat # T9424; United States) was used, Revert Aid First Strand cDNA Synthesis Kit (Thermo Scientific Cat # K1622; United States) and SYBR Green Master Mix (Applied biosystems Cat# A25742; United States) were used. Crystal violet (SRL Cat# 28376, India) was used for CV assay and SYTO9 green fluorescent nucleic acid stain (Invitrogen Cat # S34854, United States) for fluorescence microscopy.

Rathi B., Gupta S., Kumar P., Kesarwani V., Dhanda R.S., Kushwaha S.K, & Yadav M. (2022). Anti-biofilm activity of caffeine against uropathogenic E. coli is mediated by curli biogenesis. Scientific Reports, 12, 18903.

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Comparative Analysis of E. coli Strains

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A panel of Big-Six group of ATCC E. coli O111:H8, O145, O26:H11, O103:H11, O45:H2, O121:H19, and K12 MG1655 strains were purchased from ATCC. The complete strain details, genome size, and predicted protein numbers are represented in Supporting Information Table 4. All reagents were purchased from Sigma, unless otherwise specified.

Nirujogi R.S., Muthusamy B., Kim M.S., Sathe G.J., Lakshmi P.T., Kovbasnjuk O.N., Prasad T.S., Wade M, & Jabbour R.E. (2017). Secretome analysis of diarrhea-inducing strains of Escherichia coli. Proteomics, 17(6), 10.1002/pmic.201600299.

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Comparative Analysis of E. coli Strains in Ionic Liquids

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Two E. coli strains were utilized: DH1 (ATCC 33849) and K-12 MG1655 (ATCC 47076). Two ionic liquids were utilized. 1-Butyl-3-methylimidazolium chloride ([C₄C₁Im]Cl) was purchased from Sigma-Aldrich (Basionics ST 70, BASF), and 1-ethyl-3-methylimidazolium acetate, ([C₂C₁Im][OAc]), was purchased from IOLITEC ionic Liquids Technologies GmbH (Heilbronn, Germany). Chemicals and components of the medium used for selecting the best performing strains were purchased from Sigma-Aldrich (St. Louis, USA) or VWR (West Chester, USA) unless otherwise noted.
M9 glucose medium contained 2 g/L glucose, 1× M9 salts, 2 mM MgSO₄, 100 µM CaCl₂ and 1× trace elements and Wolfe’s vitamin solution. Composition of 10× M9 salts solution consisted of 68 g/L Na₂HPO₄ anhydrous, 30 g/L KH₂PO₄, 5 g/L NaCl, and 10 g/L NH₄Cl dissolved in Milli-Q filtered water and autoclaved. M9 trace elements was a 2000× solution containing of 3.0 g/L FeSO₄·7H₂O, 4.5 g/L ZnSO₄·7H₂O, 0.3 g/L CoCl₂·6H₂O, 0.4 g/L Na₂MoO₄·2H₂O, 4.5 g/L CaCl₂·H₂O, 0.2 g/L CuSO₄·2H₂O, 1.0 g/L H₃BO₃, 15 g/L disodium ethylene-diamine-tetra-acetate, 0.1 g/L KI, 0.7 g/L MnCl₂·4H₂O and concentrated HCl dissolved in Milli-Q filtered water and sterile filtered. The final concentrations of the vitamin mix and trace elements in the M9 medium were 1×.

Mohamed E.T., Wang S., Lennen R.M., Herrgård M.J., Simmons B.A., Singer S.W, & Feist A.M. (2017). Generation of a platform strain for ionic liquid tolerance using adaptive laboratory evolution. Microbial Cell Factories, 16, 204.

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