Applications Information

Procedure A: Prepare Autoclavable MEM Powdered Medium (1X) (Cat. no. 11700*)

  1. To a mixing container that is as close to the final volume as possible, add 950 mL of distilled water.
  2. Add powdered medium to room temperature (15°C to 30°C) water with gentle stirring.
  3. Rinse inside of package to remove all traces of powder.
  4. When completely dissolved, adjust the pH to 4.1 to 4.2 with 1 N HCl.
  5. Dispense 95 mL of the liquid into 100-mL bottles. Do not cap tightly.
  6. Autoclave the medium for at least 15 minutes at 121°C on a slow exhaust cycle. However, due to equipment variability, we recommend that you qualify your autoclave sterilization cycle.
  7. Allow the medium to cool to room temperature.
  8. Aseptically add 3 mL of 7.5% NaHCO3 solution to each bottle. Mix well.
  9. Aseptically add L-glutamine and antibiotic solutions, serum, or other supplements, as desired. Mix well.
  10. Aseptically adjust the medium to final pH 7.2 to 7.4 by slowly adding, with stirring, 1 N NaOH or 1 N HCl, if necessary. *For in vitro diagnostic use. Caution: Not for human or animal therapeutic use.

 

 

Procedure B: Prepare Powdered Leibovitz’s L-15 Medium (1X) (Cat. no. 41300*)

  1. To a mixing container that is as close to the final volume as possible, add 950 mL of distilled water.
  2. Add powdered medium to room temperature (15°C to 30°C) water with gentle stirring. Do not heat the water.
  3. Rinse the inside of package to remove all traces of powder.
  4. Adjust pH to 7.6 by slowly adding, with stirring, 1 N NaOH or 1 N HCl.
  5. Adjust the final volume with distilled water.
  6. Process the medium immediately into sterile containers by membrane filtration with a 0.2-μm filter using a positive pressure system. *For in vitro diagnostic use. Caution: Not for human or animal therapeutic use.

 

 

Procedure C: Prepare Powdered Iscove’s Modified Dulbecco’s Medium (1X) (Cat. nos. 42200*, 12200*)

  1. To a mixing container that is as close to the final volume as possible, add 950 mL of distilled water.
  2. Add powdered medium to room temperature (15°C to 30°C) water with gentle stirring. Do not heat the water.
  3. Rinse the inside of package to remove all traces of powder.
  4. Add 3.024 g of sodium bicarbonate/L of medium.
  5. Adjust the final volume with distilled water and stir until dissolved.
  6. Do not adjust the pH. Keep the container closed until the medium is filtered.
  7. Process the medium immediately into sterile containers by membrane filtration with a 0.2-μm filter using a positive pressure system. *For in vitro diagnostic use. Caution: Not for human or animal therapeutic use.

 

 

Procedure D: Prepare Powdered Medium

  1. To a mixing container that is as close to the final volume as possible, add 950 mL of distilled water.
  2. Add powdered medium to room temperature (15°C to 30°C) water with gentle stirring. Do not heat the water.
  3. Rinse the inside of package to remove all traces of powder.
  4. Add sodium bicarbonate (see Tables 1, 2 for quantity) to medium.
  5. Adjust the pH to 0.2 and 0.3 units below the desired final working pH by slowly adding, with stirring, 1 N NaOH or
  6. 1 N HCl. The pH may rise 0.1 to 0.3 units upon filtration.
  7. Adjust the final volume with distilled water.
  8. Process the medium immediately into sterile containers by membrane filtration with a 0.2-μm filter using a positive-pressure system.

Table 1 Powdered Media Preparation (Procedure D)

Cat. no.*NaHCO3 Recommended working pH
 g/LmL/L 7.5% soln. 
113000.354.75.9–6.2
210110.8511.357.0–7.4
236002.026.666.8–7.2

 *For research use only. Caution: Not for human or animal diagnostic or therapeutic
uses.

Table 2 Powdered Media Preparation (Procedure D) 

Cat. no.*NaHCO3 Recommended working pH
 g/LmL/L 7.5% soln. 
100120.354.77.0–7.4
110120.354.77.0–7.4
119002.229.37.0–7.4
120002.229.37.0–7.4
121003.749.37.0–7.4
124001.216.07.0–7.4
125002.43832.57.0–7.4
128003.749.37.0–7.4
130180.8511.357.0–7.4
217001.1815.77.5–7.9
221002.7536.66.7–7.1
226002.432.07.1–7.4
234002.026.77.0–7.4
311002.229.37.1–7.4
316003.749.37.0–7.4
318002.026.77.0–7.4
325002.4332.57.1–7.4
415002.229.37.0–7.4
518002.026.77.0–7.4
611002.229.37.0–7.4

*For in vitro diagnostic use. Caution: Not for human or animal therapeutic use

Table 2 Powdered Media Preparation (Procedure D) - continued

Cat. no.*NaHCO3 Recommended working pH
 g/LmL/L 7.5% soln. 
236002.026.76.8–7.2
430000.354.76.1–6.3


*For research use only. Caution: Not for human or animal diagnostic or therapeutic uses.

 

 Procedure E: Prepare Powdered Medium (Alternate Recipe)

  1. To a mixing container that is as close to the final volume as possible, add 950 mL of distilled water.
  2. Add powdered medium to room temperature (15°C to 30°C) water with gentle stirring. Do not heat the water.
  3. Rinse the inside of package to remove all traces of powder.
  4. Add sodium bicarbonate (see Table 3 for quantity) to medium.
  5. Adjust the pH to 0.2 and 0.3 units below the desired final working pH by slowly adding, with stirring, 1 N NaOH or 1 N HCl.
  6. The pH may rise 0.1 to 0.3 units upon filtration.
  7. Adjust the final volume with distilled water.
  8. Process the medium immediately into sterile containers by membrane filtration using a positive-pressure system. 

Table 3 Powdered Media Preparation (Procedure E)

Cat. no.*NaHCO3 Recommended working pH
 g/LmL/L 7.5% soln. 
100120.354.77.0–7.4
110120.354.77.0–7.4
217001.17615.77.6–7.9
325002.438157.1–7.4
424001.125157.1–7.4
521003.749.37.0–7.4

*For in vitro diagnostic use. Caution: Not for human or animal therapeutic use.


Cat. no.*NaHCO3 Recommended working pH
 g/LmL/L 7.5% soln. 
100120.354.77.0–7.4

*For research use only. Caution: Not for human or animal diagnostic or therapeutic uses.

 

 

Procedure F: Liquid Media: Prepare 1X Solutions from 10X Concentrates

To prepare an acceptable, final 1X solution, perform the following procedure under aseptic conditions. Use Distilled
Water (Cat. no. 15230) and 7.5% Sodium Bicarbonate Solution (Cat. no. 25080) for use in this protocol. Also note, because we adjust the pH of all 10X media and 10X balanced salt solution concentrates for solubility, you may need to adjust the pH after the dilution and to add sodium bicarbonate, as appropriate.

  1. Aseptically dilute 100 mL of 10X concentrate with approximately 850 mL of distilled water.
  2. Aseptically add the correct amount of 7.5% sodium bicarbonate solution. See Table 4 for concentrations.
  3. Adjust the pH as necessary with 1 N HCl or 1 N NaOH. See Table 3 for 1X pH values.
  4. Adjust the final volume with distilled water.
  5. Dispense the solution into sterile containers. Cap the bottles tightly with sterile closures and store at the recommended temperature (see product label).

Table 4 pH of the Medium after Dilution and Addition of 7.5% Sodium Bicarbonate Solution (Cat. no. 25080)

Cat. no.*  
pH of
10X soln
NaHCO3 pH after 1:10
dilution &
NaHCO3
addition a
     pH of 1X soln.a               
                      g/LmL/L 7.5% soln.  
114304.8–5.32.229.37.6–7.97.0–7.4
118252.4–2.72.229.37.6–7.97.1–7.4
21180 2.0–2.82.229.37.6–7.97.1–7.4
214304.8–5.32.229.37.6–7.97.0–7.4


a: pH range varies depending on the pH of water used in dilution. *For in vitro diagnostic use. Caution: Not for human or animal therapeutic use.