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Vegetal Media for Growth of E. Coli Cultures

Formedium has added six vegetal media to its range of culture media for growth of E. coli. In all six vegetal media, Tryptone has been replaced by Soya Peptone, for the following formulations of LB Miller medium, LB Lennox medium and 2YT medium. Tryptone and Yeast Extract are present as a general nitrogen source for growth and protein expression. Both are excellent sources for nitrogen and additionally Yeast Extract provides B-type vitamins, carbohydrates and growth factors.

In vegetal media,thus avoiding any contamination of medium with constituents of animal origin, Tryptone is replaced by Soya Peptone. In contrast to Tryptone, which is a pancreatic digest of bovine milk casein, its replacement Soya Peptone is made by Papaic hydrolysis of Soya Bean meal. Nitrogen content of Soya Peptone (9.5 %) compared to Tryptone is slightly lower and the same for its amino nitrogen content (3 %). This difference of less than two percent does not notably influence growth and protein expression.

“A comparative study between the use of Tryptone as a nitrogen source and Soya Peptone as an alternative source showed no difference in growth.”

In liquid media cell growth of E. Coli is measured by optical density at 680nm this did not show any significant variance between growth curves of cells cultured in LB Miller Broth, LB Lennox Broth and 2YT Broth medium containing Tryptone or Soya Peptone.
In solid media counts of Colony Forming Units (CFU) of LB Lennox Agar, LB Miller Agar and 2YT Agar containing Tryptone or Soya Peptone did not significantly differ from each other.

No substantial difference could be noted between media containing Soya Peptone and Tryptone. Soya Bean Peptone performed as well compared to the use of Tryptone in both liquid and solid media.

In all three media LB Miller Vegetal medium, LB Lennox Vegetal medium and 2YT Vegetal medium including Soya Peptone, no animal sourced constituents are present. Due to the absence of animal sourced components Formedium certifies;

  • No raw material sourced or derived from animals are used in the manufacture of this product.
  • The materials used in the production are in compliance with the European directive 75 / 318 / EEC as amended by Directive 1999 / 82 / EC.

Therefore Formedium Ltd. takes the position that this product is free of any risk in terms of Bovine Spongiform Encephalothy (BSE) or Transmissible Spongiform Encephalopathy (TSE).

Formedium Vegetal media

Methods:
Broth culture: Cell growth was measured in LB Lennox, LB Miller and 2YT medium.
Each medium was present in a formulation containing Tryptone or Soya Peptone.

  • Single colony incubated in 5 ml of all three media tested (3 Tryptone vs 3 Soya Bean peptone) O/N @ 37 °C 180 rpm.
  • Following day 1:100 dilution, 30 ml culture in 150 ml flask incubated @ 37 °C 180 rpm. Growth measured by optical density at 680nm, measured hourly
  • 3 biological repeats
  • LB Lennox (g/l); Soya peptone 10 or Tryptone 10, Yeast extract 5, NaCl 5.
    LB Miller (g/l); Soya peptone 10 or Tryptone 10, Yeast extract 5, NaCl 10.
    2YT; (g/l); Soya peptone 16 or Tryptone 16, Yeast extract 10, NaCl 5.

Formedium Vegetal media

Methods:
Agar culture on plates: The number of Colony Forming units was measured in LB Lennox, LB Miller and 2YT Agar plates.
Each medium was present in a formulation containing Tryptone or Soya Peptone.

  • Single colony incubated in 5 ml of all three media tested (3 Tryptone vs 3 Soya Bean peptone) O/N @ 37 °C 180 rpm.
  • Following day 100 µl of 10-fold dilution series (to 10-6) was spread onto the same medium as agar (e.g., Miller broth colony onto Miller agar, Lennox-Soya colony onto Lennox-Soya agar). Incubated O/N @ 37 °C
  • Colony count (cfu/ml) for 3 biological repeats
  • LB Lennox (g/l); Soya peptone 10 or Tryptone 10, Yeast extract 5, NaCl 5, Agar 15.
    LB Miller (g/l); Soya peptone 10 or Tryptone 10, Yeast extract 5, NaCl 10, Agar 15.
    2YT; (g/l); Soya peptone 16 or Tryptone 16, Yeast extract 10, NaCl 5, Agar 15.
  • Soya Peptone

    $31.68$528.00
    SKU Pack Size Price
    VPEP01 250g $31.68
    VPEP02 500g $55.11
    VPEP03 1000g $95.70
    VPEP04 6 x 1kg $528.00
    Soya Peptone is a papaic digest of defatted soybean flour and is a well-balanced source of essential amino acids, carbohydrates and vitamins in cell cultures. Soya Peptone is used for growth of a wide variety of bacteria and yeasts in cell cultures and is often combined with Tryptone or Peptone for a rapid and abundant growth of cells. This plant peptone is classified animal-free by Formedium Ltd. Based on the manufacturing protocol, we attest that no animal raw materials are prescribed for use in the production of this product, nor are any of the raw materials derived from animal products. Soya Peptone is classified animal free, GMO free (according to the European Directive 2001/18/CE) Store dry at room temperature
  • VLB-Agar Lennox

    $4.60$330.20
    SKU Pack Size Price
    VLBAL1L 1 Litre pack $4.60
    VLBAL2L 2 Litre pack $6.98
    VLBAL5L 5 Litre pack $13.60
    VLBAL10L 10L pack $23.15
    VLBAL20L 20 Litre pack $43.35
    VLBAL0250 250g $18.00
    VLBAL0500 500g $35.64
    VLBAL1000 1000g $61.35
    VLBAL6000 6 x 1kg $330.20
  • VLB-Broth Lennox

    $4.22$276.21
    SKU Pack Size Price
    VLBBL1L 1 Litre pack $4.22
    VLBBL2L-1 2 Litre pack $6.80
    VLBBL5L 5 Litre pack $9.74
    VLBBL10L 10L pack $16.90
    VLBBL20L 20 Litre pack $27.01
    VLBBL0250 250g $17.82
    VLBBL0500 500g $32.32
    VLBBL1000 1kg $58.95
    VLBBL6000 6 x 1kg $276.21
  • VLB-Agar Miller

    $5.33$438.19
    SKU Pack Size Price
    VLBAM1L 1 Litre pack $5.33
    VLBAM2L 2 Litre pack $8.09
    VLBAM5L 5 Litre pack $18.93
    VLBAM10L 10L pack $34.72
    VLBAM20L 20 Litre pack $65.57
    VLBAM0250 250g $25.54
    VLBAM0500 500g $46.66
    VLBAM1000 1000g $81.00
    VLBAM6000 6 x 1kg $438.19
  • VLB-Broth Miller

    $4.22$306.70
    SKU Pack Size Price
    VLBBM1L 1 Litre pack - 25g $4.22
    VLBBM2L 2 Litre pack - 50g $5.89
    VLBBM5L 5 Litre pack - 125 gram $11.58
    VLBBM10L 10 Litre pack - 250g $19.65
    VLBBM20L 20 Litre pack - 500g $36.73
    VLBBM0250 250g $19.65
    VLBBM0500 500g $36.73
    VLBBM1000 1000g $60.98
    VLBBM1500 1500g $86.87
    VLBBM6000 6 x 1kg $306.70
     
  • 2X VYT Broth

    $5.33$404.02
    SKU Pack Size Price
    VYTB1L 1 Litre pack - 31g $5.33
    VYTB2L 2 Litre pack $8.09
    VYTB5L 5 Litre pack $13.60
    VYTB10L 10 Litre pack - 310g $26.09
    VYTB20L 20 Litre pack $50.14
    VYTB0250 250g $21.32
    VYTB1000 1000g $73.84
    VYTB6000 6 x 1kg $404.02
  • 2X VYT Agar

    $6.98$505.58
    SKU Pack Size Price
    VYTA1L 1 Litre pack $6.98
    VYTA2L 2 Litre pack $10.48
    VYTA5L 5 Litre pack $24.26
    VYTA10L 10L pack $46.28
    VYTA20L 20 Litre pack $88.70
    VYTA0250 250g $26.27
    VYTA1000 1000g $93.49
    VYTA6000 6 x 1kg $505.58