Instructions for preparation of single-strength medium

Directions for the preparation of single strength synthetic liquid media (1X) from concentrated media


  • Measure out sterile culture-grade water (Cat. # 03-055-1) to approximately 70% of desired total volume of medium. Pour water into an appropriate sterile mixing container that is close to the desired final volume. The water should be at room temperature.
  • Add the amount of the concentrated medium.
  • Add the desired amount of L-Glutamine Solution 200mM (Cat. # 03-020-1) if required (See table)*.
  • Add the desired amount of Sodium Bicarbonate Solution (Cat. # 03-040-1). (See table).
  • Add antibiotics solution if desired*.
  • Add water to the final volume. During the dilution, mix gently to obtain a uniform solution. If necessary, adjust pH with sterile 1N NaOH or HCl.
  • Store at 2ºC to 8ºC.

* To avoid shortening the shelf life, add these components just before use.

Important: The above procedures are carried out under strict sterile environment.

Example I, without sterile filatration (asceptic technique)

Preparation of Basal Medium Eagle, Earle’s Salts Base, one liter:

  • 700 ml sterile water (Cat. # 03-055-1)
  • 100 ml Basal Medium Eagle (Earle's) 10X w/o L-Glutamine, w/o Sodium Bicarbonate (Cat. # 01-015-5).
  • 10 ml L-Glutamine Solution 200mM (Cat. # 03-020-1)
  • 29.4 ml Sodium Bicarbonate Solution 7.5% (Cat. # 03-040-1).
  • 10 ml Penicillin-Streptomycin Solution (Cat. # 03-031-1)
  • Adjust pH if necessary with sterile 1N NaOH or 1N HCl. Sterile water to final volume.
  • Example II, with sterile filtration (non-asceptic technique)

Example II, with sterile filtration (non-asceptic technique)

Preparation of RPMI 1640, 1 liter:

  • 700 ml distilled water
  • Add 100 ml RPMI concentrate 10X
  • Adjust pH to 6.5-7.0 with 1N NaOH
  • Add 10.3 ml L-Glutamine Solution 200mM
  • Add 26.7 ml Sodium Bicarbonate Solution 7.5%
  • Adjust pH with 1N NaOH or 1N HCl to pH 7.0-7.4
  • Add distilled water to final volume. Adjust pH if necessary.
  • Filter the medium into sterile container using a 0.2µm membrane filter.

Recommended amounts of Sodium Bicarbonate and L-Glutamine to be added in the preparation of single strength liquid media (1X) from concentrated media (5X or 10X)

Product DescriptionDesired Cat. No.Prepared from Cat. No.Sodium Bicarbonate Solution 7.5%,L-Glutamine Solution 200mM,
      Cat.# 03-040-1 Cat.# 03-020-1
      ml/Liter ml/liter
Basal Medium-Eagle Earle’s salts base 01-015-1 01-015-5 29.4 10
         
Minimum Essential Medium Eagle Earle’s Salts Base 01-025-1 01-025-5 29.4 10
         
Dulbecco’s Modified Eagle Medium (DMEM), Low Glucose 01-050-1 01-050-4 (5X) 49.4 20
         
Dulbecco’s Modified Eagle Medium (DMEM), High Glucose 01-055-1 01-055-4 (5X) 49.4 20
         
Glasgow Modified MEM*(GMEM) 01-060-1 01-060-5 36.7 20
         
Medium M-199 Earle’s Salts Base 01-080-1 01-080-5 29.4  
         
Medium M-199 Hanks’ Salts Base 01-085-1 01-085-5 4.7  
         
Nutrient Mixture F10 (HAM) 01-090-1 01-090-5 16  
         
Nutrient Mixture F12 (HAM) 01-095-1 01-095-5 15.7  
         
RPMI 1640 01-100-1 01-104-5 26.7  
        10.3
DMEM F12 (1:1) 01-170-1 01-170-5 16  
        12.5
DCCM-1 05-010-1 05-010-5 29.4 10-20
         
DCCM-2 05-015-1 05-015-5 29.4 10-20
         
LPM 05-040-1 05-040-5 29.4 10-20
         
BIO-MPM-1 05-060-1 05-060-5 26.9 10-20
         
BIOCHO-1 SFM Base 05-061-1 05-061-5 26.9 10-20
         
BIOCHO-2 SFM Base 05-062-1 05-062-5 4.7 10-20