ATCC Medium: 1034 Modified PYNFH Medium - IBEMicrobeHub/culturing GitHub Wiki
Materials:
- Filters 0,22 µm.
- Aliquots.
- HCl 37%.
| Medium (table 1) | Quantity |
|---|---|
| Peptone (Ref. 211677, BD) | 10 g |
| Yeast Extract (Ref. 212750, BD) | 10 g |
| Yeast nucleic acid (Ribonucleic Acid, Type VI from Torula Yeast) (Sigma#R-6625) | 1 g |
| Folic acid (Sigma#F8758) | 0,015 g |
| Distilled water | 880 mL |
| Components (table 2) | Quantity |
|---|---|
| Hemin solution (Sigma#H9039) (table 3) | 0,4 mL |
| Buffer solution (table 4) | 20 mL |
| Fetal bovine serum (heat-inactivated)(Sigma#F9665-100ml) | 100 mL |
| Hemin solution (table 3) | Quantity |
|---|---|
| NaOH | 0,4 g |
| Distilled water | 20 mL |
| Hemin (Sigma#H9039) | 0,5 g |
| Buffer solution (table 4) | Quantity |
|---|---|
| KH2PO4 (Sigma#P5655) | 18,1 g |
| Na2HPO4 (Sigma#S5136) | 25 g |
| Distilled water | 1 L |
Method:
- Prepare the Hemin solution (table 3):
- Dissolve 0,4 g of NaOH in a 200 mL of distilled water.
- Add 0,5 g of Hemin.
- Autoclave 20 min at 121ºC.
- Dispense 0,4 mL of the solution into aliquots.
- Store at 4ºC and protected from the light.
- Prepare the buffer solution (table 4).
- Prepare the medium (table 1).
- Autoclave for 15 min at 121ºC.
- Let it cool and aseptically add the components of table 2.
- Adjust the final pH with HCl 37% and filter.
- Store at 4ºC.
Culture list:
- Capsaspora owczarzaki: 23ºC.
- Naegleria gruberi: 29ºC.