| 2HPO4•3H2O | 28 g |
| K2HPO4, anhydrous | 21.4 g |
| Milli-Q Water | 500 mL |
- Measure out Milli-Q
- Add either of two chemicals
- Autoclave both containers
- Store at 4oC
Media
-Pi Stock
| MgSO4•7H2O | 20 g |
| CaCl2•2H2O | 6 g |
| NaCl | 20 g |
| Microelement Stock | 500 mL |
| Milli-Q Water | 1.5 L |
- Dilute first three ingredients separately into 500 mL of Milli-Q
- Autoclave three solutions, and allow to cool
- Combine four solutions in order, allowing each to dissolve
- Store at 4oC, shake well before use, do not autoclave
PGY
| Yeast Extract | 250 mg |
| Peptone | 250 mg |
| Glucose | 250 mg |
| Agar | 15 g |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Add all components in order
- Bring volume up to 1 L with Milli-Q
- Autoclave for 20 minutes
PES30 (Agar)
| Instant Ocean | 30 g |
| PES Stock | 20 mL |
| Vitamin Solution | 2 mL |
| (Agar | 10 g) |
| Milli-Q Water | ~1 L |
- Measure out Milli-Q
- Add and dissolve Instant Ocean
- Lower pH to approximately 3
- Bubble overnight
- Raise pH to 8.3
- (Add agar)
- Autoclave
- Add PES stock and vitamin solution
- Aliquot and store as needed
pH: 8.3
MN
| NaNO3 | 750 mg |
| K2HPO4•3H2O | 20 mg |
| MgSO4•7H2O | 38 mg |
| CaCl2•2H2O | 18 mg |
| Citric Acid | 3 mg |
| Ferric Ammonium Citrate | 3 mg |
| EDTA (disodium magnesium) | 0.5 mg |
| Na2CO3 | 20 mg |
| Trace Metal Solution | 1 mL |
| Sea Water | 750 mL |
| Milli-Q Water | ~250 mL |
- Measure out Sea Water
- Add all components in order
- Bring volume up to 1 L with Milli-Q
- Autoclave for 20 minutes
pH: 8.3
IMR Saltwater Medium
| NaNO3 (4.25 g/L) | 10 mL |
| NaH2PO4 (690 mg/L) | 7 mL |
| Trace Metal Stock | 1 mL |
| Vitamin Stock | 500 µL |
| Sea Water (filtered) | 1 L |
- Measure out sea water
- Add first three components in order
- Bring volume up to 1 L with Milli-Q
- (Optional: Add 10 g Gelrite for solid medium)
- Autoclave for 20 minutes
- Add vitamin stock
- Store at 4oC
D Medium
Stock
| NTA | 2 g |
| NaNO3 | 14 g |
| KNO3 | 2 g |
| Na2HPO4 | 2.2 g |
| MgSO4•7H2O | 2 g |
| CaSO4•2H2O | 1.2 g |
| NaCl | 160 mg |
| Iron Solution | 20 mg |
| Micronutrient Solution | 10 mL |
| Milli-Q Water | ~1 L |
- Measure out ~500 mL Milli-Q Water
- Add components in order
- Bring volume to 1 L with Milli-Q
- Adjust pH to 8.2
- Autoclave for 20 minutes
Media
| D Medium Stock | 50 mL |
| Milli-Q Water | ~1 L |
- Measure out ~500 mL Milli-Q
- Add D Medium Stock
- Bring volume to 1 L with Milli-Q
- Adjust pH to 8.2
- Either add 1% agar, autoclave for 20 minutes and pour plates, or aliquote into erlenmeyer flasks and autoclave for 20 minutes
BG11o-PGY
| BG11o | 100 mL |
| Yeast Extract | 250 mg |
| Peptone | 250 mg |
| Glucose | 250 mg |
| Agar | 15 g |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Add all components in order
- Bring volume up to 1 L with Milli-Q
- Adjust pH to 7.4
- Autoclave for 20 minutes
pH: 7.4
BG11-PGY
| BG-11 | 100 mL |
| Yeast Extract | 250 mg |
| Peptone | 250 mg |
| Glucose | 250 mg |
| Agar | 15 g |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Add all components in order
- Bring volume up to 1 L with Milli-Q
- Adjust pH to 7.4
- Autoclave for 20 minutes
pH: 7.4
BG11o
| Na2-EDTA | 1 mg |
| Citric Acid | 6 mg |
| K2HPO4•3H2O | 40 mg |
| MgSO4•7H2O | 75 mg |
| CaCl2•2H2O | 36 mg |
| Na2CO3 (H2O) | 20 mg |
| Ferric Ammonium Citrate | 6 mg |
| Trace Metal Solution | 50 µL |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Add all components in order
- Bring volume up to 1 L with Milli-Q
- Autoclave for 20 minutes
(note: Do Not Autoclave if making concentrated stock solutions!)
BG11
| NaNO3 | 1.5 g |
| K2HPO4•3H2O | 40 mg |
| MgSO4•7H2O | 75 mg |
| CaCl2•2H2O | 36 mg |
| Citric Acid | 6 mg |
| Ferric Ammonium Citrate | 6 mg |
| EDTA (disodium magnesium) | 1 mg |
| Na2CO3 | 20 mg |
| Trace Metal Solution | 1 mL |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Add all components in order
- Bring volume up to 1 L with Milli-Q
- Autoclave for 20 minutes
(note: Do Not Autoclave if making concentrated stock solutions!)
pH: 7.4
ATCC Culture Medium 929
| Ammonium Sulfate | 1.32 mg |
| MgSO4•7H2O | 380 mg |
| CaCl2•2H2O | 20 mg |
| MnCl2•4H2O, 50.5 mM solution | 20 µL |
| Na2MoO4•2H2O, 41.3 mM solution | 10 µL |
| CoCl2•6H2O, 1.7 mM solution | 5 µL |
| ZnSO4•7H2O, 34.8 mM solution | 10 µL |
| K2HPO4 | 87 mg |
| Phenol red 0.5% | 250 µL |
| Iron Solution | 250 µL |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Add all components in order
- Bring volume up to 1 L with Milli-Q
- Adjust pH to 7.5 with 0.5M K2CO3
- Autoclave for 20 minutes
pH: 7.5
ASNIII
| NaCl | 25 g |
| MgCl2•6H2O | 2 g |
| KCl | 500 mg |
| NaNO3 | 750 mg |
| K2HPO4•3H2O | 20 mg |
| MgSO4•7H2O | 3.5 g |
| CaCl2•2H2O | 500 mg |
| Citric Acid | 3 mg |
| Ferric Ammonium Citrate | 3 mg |
| EDTA (disodium magnesium) | 0.5 mg |
| Na2CO3 | 20 mg |
| Trace Metal Solution | 1 mL |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Add all components in order
- Bring volume up to 1 L with Milli-Q
- Autoclave for 20 minutes
pH: 7.5
Allen and Arnon Medium (Plates)
| -Pi stock | 25 mL |
| +Pi stock | 6.25 mL |
| Agar | 10 g |
| Streptomycin Stock (1 mg/mL) | 1 mL |
| Neomycin Stock (25 mg/mL) | 1 mL |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Shake -Pi stock
- Add both stocks to water
- Bring volume up to 1 L with Milli-Q
- Check pH. If less than 7.8, make fresh +Pi
- Add agar
- Autoclave for 20 minutes
- Add antibiotics
Allen and Arnon Medium (Liquid)
| -Pi stock | 6.30 mL |
| +Pi stock | 3.10 mL |
| Streptomycin Stock (1 mg/mL) | 1 mL |
| Neomycin Stock (25 mg/mL) | 1 mL |
| Milli-Q Water | ~1 L |
- Measure out approximately 300 mL Milli-Q
- Shake -Pi stock
- Add both stocks to water
- Bring volume up to 1 L with Milli-Q
- Check pH. If less than 7.8, make fresh +Pi
- Autoclave for 20 minutes
- Add antibiotics