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Grace's MediumPowdered Ham's F-12 Nutrient Mixture

Powdered Ham’s F-12 Nutrient Mixture

Ham’s F-12 Nutrient Mixture is a cell culture medium that was designed by Ham in 1969 based on Ham’s F-10 for serum-free culture of CHO cells.


Ham’s F-12 Nutrient Mixture contains a wider range of components than other basal media, including zinc, putrescine, hypoxanthine and thymidine. It is often used as a basal medium for serum-free culture and is particularly suitable for single-cell culture and cloning under low serum conditions. It is also widely used for the culture of cancer cells and primary cells such as rat liver cells, rat prostate epithelial cells, chondrocytes, rat muscle cells, chicken embryo cells and more. Ham’s F-12 does not contain protein, lipids or growth factors. Therefore, it may need to be supplemented with nutrients and usually requires the addition of 10% fetal bovine serum (FBS). Ham’s F-12 uses a bicarbonate buffer system and therefore requires a 5-10% CO2 environment to maintain physiological pH value.
We offer various modified Ham’s F-12 media formulations with different components for a wide range of cell culture applications.

Ingredients of Powdered Ham’s F-12 Nutrient Mixture

P/NSpecificationIngredient Description
HCY-BCMP09001A1L×10 bagIncluding: L-glutamine, phenol red
Not contain: sodium bicarbonate
HCY-BCMP09001B10L×1 bottleIncluding: L-glutamine, phenol red
Not contain: sodium bicarbonate

This Ham’s F-12 dry powder cell culture medium needs to be supplemented with sodium bicarbonate, pH adjusted and filtered during preparation. The detailed preparation plan is as follows:

  1. Add 950 mL of distilled water to the mixing container as close as possible to the final volume.
  2. Add the dry culture medium to room temperature (15°C to 30°C) water and stir gently. Do not heat the water.
  3. Rinse the inside of the package to remove all residual powder.
  4. Add 15.7mL of 75g/L NaHCO3 solution to the medium.
  5. Adjust the pH value to the desired final working pH below 0.2 to 0.3 units by slowly adding and stirring 1 N NaOH or 1 N HCl. The pH value may increase by 0.1 to 0.3 units after filtration. The recommended working pH value is 7.5-7.9.
  6. Adjust the final volume with distilled water.
  7. Immediately use a positive pressure filtration system to filter the medium through a 0.2 μm filter membrane into a sterile container.

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