Media for Optimal Cell Growth and Yield of
Escherichia Coli Cultures
LB Media (Luria-Bertani) are common bacterial growth media for Escherichia Coli.
Although already described in the fifties in the early days of phage genetics these media are still widely used in molecular biology.
The two main components of LB media are Tryptone and Yeast Extract. Tryptone is used in a concentration of 10 gram / litre and Yeast Extract in a concentration of 5 gram / litre. Many variations of LB medium only differs in the concentration of NaCl. All LB-Media are listed in order of increasing concentration of NaCl.
LB Medium Miller is the High Salt LB medium type with 10 gram /litre whereas LB Medium Lennox is the Low Salt LB medium type with 5 gram / litre. L-Broth only contain 0.5 gram of NaCl per litre and is used mainly when working with phage P1 where CaCl2 is added for efficient adherence of the phage to the cell. In LBM media Mg2+ is added to enhance adsorption of phage lambda to the cells. Tryptone and Yeast Extract do not contain enough Mg2+ for optimal adsorption.
Many synonymous names are know for several different type of LB media. We have tried to rank them all.
In case a medium cannot be found please also look at the synonymous names.
Tryptone broth is a moderately rich medium for growth and cultivation of Escherichia Coli.
Terrific Broth is a rich medium compared to LB and Tryptone Media. The medium is developed for higher density growth of Escherichia Coli cells and higher yield of plasmid DNA compared to LB and Tryptone broth.
Super Broth is an even richer medium developed for obtaining high yields of lambda bacteriophage in liquid lysates, Botstein, D. et al.
Powdered Media Storage and Preparation
Ready made powdered media are hygroscopic and must be protected from atmospheric moisture. Always reseal tightly after opening.
Store the medium dry at room temperature.
Preparing media in a concentrated form is not recommended. Some salt and protein complexes may precipitate in a concentrated solution.
Sterilise the medium in a validated autoclave at 1 kg/cm2 (15 psi) at 121°C. for 15 minutes. Higher temperatures or exceeding the period of time for autoclavation may cause a brown decolourisation of the medium due to caramelisation of the glucose present in the medium. Denaturising of proteins may occur as well. Both will result in poor cell growth.
References:
Botstein, D. et al, Mol. Biol., 91, 439, (1975)
Lennox, E.S., Virology, 1, 190, (1955).
Luria, S.E. and Burrous, J.W., J. Bacteriol. 74, 471, (1957).
Luria, S.E. et al., Virology, 12, 348, (1960).
Blattner, F., et al. Science, 196, 161, (1977).
Miller, J.H., Experiments in Molecular Genetics, C.S.H. Press, N.Y., (1972)
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SKU Size Price SDA0250 250g $23.10 SDA0500 500g $42.90 SDA1000 1kg $79.20 Sabouraud Dextrose Agar (SDA) is used for the isolation, cultivation, and maintenance of non-pathogenic and pathogenic species of fungi and yeasts. SDA was formulated by Sabouraud in 1892 for culturing dermatophytes. The pH is adjusted to approximately 5.6 in order to enhance the growth of fungi, especially dermatophytes, and to slightly inhibit bacterial growth in clinical specimens. Peptones provide the nitrogen and vitamin source required for organism growth in SDA. Dextrose is added as the energy and carbon source. Agar is the solidifying agent. Store dry at room temperature -
SKU Size Price LBL0101 250g $29.83 LBL0102 1000g $78.61 LBL0103 6 x 1kg $433.22 Luria Broth Base (Miller's Modification)
Luria Broth Base or L-Broth is based upon the Luria Broth formulation as originally developed by Miller for cultivation and maintenance of E. coli cells in molecular biology. L-Broth is a nutritionally rich medium originally developed for growth and maintenance of recombinant E. coli strains. E. coli is grown to late log phase in LB Broth. These strains are generally derived from E. coli K12 which are deficient in B vitamin production. K12 has been ultimately modified by specific mutation into an auxothropic strain not capable of growth on nutritionally deficient medium. All nutritional requirements of E. coli strains are provided by LB Broth. Peptides and amino acids are abundantly present in Tryptone. Yeast extract is a rich source of amino acids, vitamins, nucleotides and carbohydrates. These nutritional elements in LB media, which otherwise the cell would have to synthesize, support a luxurious growth of E. coli cells. Sodium ions for transport and osmotic balance are provided by Sodium chloride. The concentration of NaCl in L-Broth is low compared to both LB Miller and LB Lennox formulations, respectively 10% and 5% of the NaCl concentration is present in both formulations. These variations in Sodium chloride content make it possible to select the optimal salt concentration for a specific strain. Miller, J.H., Experiments in molecular genetics, Cold Spring harbour Laboratory, Cold Spring harbour, New York, (1972). Lennox, E.S., Transuction of linked genetic characters of the host by bacteripphage P1, 1, 190-206, (1955). Sambrook, J,., E. F. Fritsch, and T. Maniatis, 1989, Molecular cloning: a laboratory manual, 2nd edition ed., Cold Spring Harbour laboratory, Cold Spring Harbour, N.Y. -
SKU Size Price LBX1L 1 Litre pack $5.28 LBX2L 2 Litre pack $8.46 LBX5L 5 Litre pack $11.98 LBX10L 10L pack $20.94 LBX20L 20 Litre pack $33.48 LBX0101 250g $22.75 LBX0104 500g $41.18 LBX0102 1kg $75.80 LBX0103 6 x 1kg $355.08 LB-Broth Low Salt, Lennox L Broth
LB Broth Lennox is a nutritionally rich medium originally developed for growth and maintenance of recombinant E. coli strains. E. coli is grown to late log phase in LB Broth. These strains are generally derived from E. coli K12 which are deficient in B vitamin production. K12 has been ultimately modified by specific mutation into an auxothropic strain not capable of growth on nutritionally deficient medium. All nutritional requirements of E. coli strains are provided by LB Broth. Peptides and amino acids are abundantly present in Tryptone. Yeast extract is a rich source of amino acids, vitamins, nucleotides and carbohydrates. These nutritional elements in LB media, which otherwise the cell would have to synthesize, support a luxurious growth of E. coli cells. Sodium ions for transport and osmotic balance are provided by Sodium chloride. The concentration of NaCl in LB Lennox is half the concentration of the LB Miller Broth formulation and ten times the concentration of LB Luria Broth formulation. These variations in Sodium chloride content make it possible to select the optimal salt concentration for a specific strain. Lennox, E.S., Transuction of linked genetic characters of the host by bacteripphage P1, 1, 190-206, (1955). Assubel, F.M., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith and K. Struhl, Current protocols in molecular biology, vol. 1, Current Protocols, New York, (1994). -
SKU Size Price VLBBM1L 1 Litre pack - 25g $5.26 VLBBM2L 2 Litre pack - 50g $7.33 VLBBM5L 5 Litre pack - 125 gram $14.37 VLBBM10L 10 Litre pack - 250g $24.35 VLBBM20L 20 Litre pack - 500g $45.51 VLBBM0250 250g $24.35 VLBBM0500 500g $45.51 VLBBM1000 1000g $75.54 VLBBM1500 1500g $107.58 VLBBM6000 6 x 1kg $379.81 -
SKU Size Price LXM0101 250g $21.19 LXM0102 1000g $70.97 LXM0103 6 x 1kg $332.03 LBM-Broth Low Salt
For propagation of of lambda phages in Escherichia coli. In LBM media Mg2 is added to enhance adsorbtion of phage lambda to the cells. Tryptone and Yeast extract do not contain enough Mg2 for optimal adsorption. -
SKU Size Price LXM0201 250g $21.65 LXM0202 1000g $73.49 LXM0203 6 x 1kg $396.86 LBM-Agar Low Salt
For propagation of of lambda phages in Escherichia coli. In LBM media Mg2 is added to enhance adsorbtion of phage lambda to the cells. Tryptone and Yeast extract do not contain enough Mg2 for optimal adsorption. -
SKU Size Price LMM1L 1 Litre pack - 25g $5.28 LMM2L 2 Litre pack - 50g $7.33 LMM5L 5 Litre pack - 125 gram $14.37 LMM10L 10 Litre pack - 250g $24.37 LMM20L 20 Litre pack - 500g $45.51 LMM0101 250g $24.37 LMM0104 500g $45.51 LMM0102 1000g $77.96 LMM0105 1500g $113.31 LMM0103 6 x 1kg $431.39 LMM0125 12.5kg $827.66 LB-Broth High Salt, Miller's LB Broth, Luria Broth
LB Broth Miller is a nutritionally rich medium originally developed for growth and maintenance of recombinant E. coli strains. E. coli is grown to late log phase in LB Broth. These strains are generally derived from E. coli K12 which are deficient in B vitamin production. K12 has been ultimately modified by specific mutation into an auxothropic strain not capable of growth on nutritionally deficient medium. All nutritional requirements of E. coli strains are provided by LB Broth. Peptides and amino acids are abundantly present in Tryptone. Yeast extract is a rich source of amino acids, vitamins, nucleotides and carbohydrates. These nutritional elements in LB media, which otherwise the cell would have to synthesize, support a luxurious growth of E. coli cells. Sodium ions for transport and osmotic balance are provided by Sodium chloride. The concentration of NaCl in LB Miller is twice the concentration of the LB Lennox Broth formulation and twenty times the concentration of LB Luria Broth formulation. These variations in Sodium chloride content make it possible to select the optimal salt concentration for a specific strain. Luria, S.E. and J.W. Burrous, Hybridization between Escherich coli and Shigella, J. Bacteriol., 74, 461-476. 1955. Luria, S.E., J.N. Adams and R.C. Ting, Transduction of lactose-utilizing ability amongst strains of E. coli and S. dysenteria and the properties of transducing phage particles, Virology, 12, 348-390, 1960. Lennox, E.S., Transuction of linked genetic characters of the host by bacteripphage P1, 1, 190-206, (1955). Sambrook, J,., E. F. Fritsch, and T. Maniatis, 1989, Molecular cloning: a laboratory manual, 2nd edition ed., Cold Spring Harbour laboratory, Cold Spring Harbour, N.Y. Assubel, F.M., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith and K. Struhl, Current protocols in molecular biology, vol. 1, Current Protocols, New York, (1994). -
SKU Size Price TRB0101 250g $25.31 TRB0104 500g $45.62 TRB0102 1000g $92.42 TRB0103 6 x 1kg $494.82 Terrific Broth is rich nutritional medium developed by Tartoff and Hobbs supporting luxurious high density growth of E.coli and a higher yield of plasmid DNA compared to LB type media. Recombinant E. coli cells have an extended growth phase in Terrific medium. Terrific Broth medium is used with glycerol as a carbon source. Unlike glucose, glycerol is not fermented to acetic acid. Tryptone and Yeast are present in the medium in elevated concentrations and provide nitrogen, vitamins and cofactors for a luxurious growth. Potassium phosphate is present in Terrific broth buffered to balance the pH and subsequently prevent cell death due to a drop in pH. Tartoff, K. D., and C. A. Hobbs, Improved media for growing plasmids and cosmid clones, Bethesda Research Laboratories, 9, 12, (1987). Sambrook, J,., E. F. Fritsch, and T. Maniatis, 1989, Molecular cloning: a laboratory manual, 2nd edition ed., Cold Spring Harbour laboratory, Cold Spring Harbour, N.Y. -
SKU Size Price SPB0101 250g $29.58 SPB0102 1000g $108.26 SPB0103 6 x 1kg $584.03 Super broth is an extremely rich medium with highly elevated levels of Tryptone and Yeast extract. Due to the high levels of peptides, amino acids, vitamins and cofactors high yields of E. coli cells and lambda bacteriophages are obtained. -
SKU Size Price MMS0101 250g $43.73 MMS0102 1000g $140.84 MMS0103 6 x 1kg $754.68 M9 Minimal Salts Base, 5x is used in preparing M9 Minimal medium used for the cultivation and maintenance of E. coli in molecular biology. M9 is a chemical defined minimal medium supporting the growth of "wild-type" strains of E.coli. The medium can be supplemented with specific amino acids or other required nutrients allowing for selection of specific auxotrophs. Glucose may be added as a source of carbohydrate. Nitrogen is provided by ammonium chloride final osmolarity of the medium is maintained by NaCl. Addition of magnesium and calcium increases the growth of the recombinant cells. M9 Minimal Salts Base, 5x is a 5x concentrate and has to be diluted to a 1x concentration and subsequently supplemented with an appropriate carbon source, such as glucose. Addition of required nutritionally elements may be added. Difco Manual, 11th edition, Sparks, MD, 272, 1998. Davis, L.G., M.D. Dibner and J.F. Battey, Basic methods in molecular biology, Elsevier, new York, (1986). Sambrook, J,., E. F. Fritsch, and T. Maniatis, 1989, Molecular cloning: a laboratory manual, 2nd edition ed., Cold Spring Harbour laboratory, Cold Spring Harbour, N.Y. -
SKU Size Price M170110 1kg $188.78 M170160 6 x 1kg $1,031.58 M17 Medium is a well buffered enriched medium for bacterial growth. The medium is optimized for isolating lactic Streptococci. Casein hydrolysate, Beef extract and Soya peptone supply carbon sources, nitrogen, vitamins and minerals essential for vigorous growth. Yeast extract supplies B-complex vitamins which stimulate bacteria growth. The medium is buffered by Disodium-β-glycerophosphate which prevents a drop in pH by the production of organic acids, such as lactic acid, during fermentation. Ascorbic acid and Magnesium sulphate are added to stimulate growth.