Media Recipies

Allen and Arnon (Liquid)	Allen and Arnon (Plates)	ASNIII	ATCC Culture Medium 929	BG11
BG11^o	BG11-PGY	BG11^o-PGY	D Medium	IMR Saltwater Medium
MN	PES30 Agar	PGY

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

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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

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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

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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

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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

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Back BG11-PGY

BG11^o

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!)

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Back To BG11^o-PGY

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

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BG11^o-PGY

BG11^o	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

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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

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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 4^oC

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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

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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

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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

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Media Recipies

6.30 mL

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

25 mL

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

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

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

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

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

NaNO3

1.5 g

K2HPO4•3H2O

40 mg

MgSO4•7H2O

75 mg

CaCl2•2H2O

36 mg

Citric Acid

6 mg

Ferric Ammonium Citrate

-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

-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

MgCl₂•6H₂O

NaNO₃

K₂HPO₄•3H₂O

MgSO₄•7H₂O

CaCl₂•2H₂O

Na₂CO₃

-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

MgSO₄•7H₂O

CaCl₂•2H₂O

MnCl₂•4H₂O, 50.5 mM solution

Na₂MoO₄•2H₂O, 41.3 mM solution

CoCl₂•6H₂O, 1.7 mM solution

ZnSO₄•7H₂O, 34.8 mM solution

K₂HPO₄

-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

NaNO₃

K₂HPO₄•3H₂O

MgSO₄•7H₂O

CaCl2•2H₂O

Na₂CO₃

-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!)

Na₂-EDTA

K₂HPO₄•3H₂O

MgSO₄•7H₂O

CaCl₂•2H₂O

Na₂CO₃ (H₂O)

-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!)

-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