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

 

 

-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

 

Microelement Stock

Fe-EDTA solution

80 mL

MnCl2•4H2O

180 mg

MoO3 (85% purity)
or
Na2Mo4•2H2O (99% purity)

18 mg
 
30.55 mg

ZnSO4•7H2O

22.0 mg

CuSO4•5H2O

7.9 mg

H3BO3 (Boric Acid)

286 mg

Vanadyl sufate hydrate

6.6 mg

CoC12•6H2O

4.0 mg

Milli-Q Water

~545 mL

 

-Measure out Milli-Q
-Dissolve all components in order
(Note: Do Not autoclave)

 

Fe-EDTA (A&A)

Na2EDTA•2H2O

866.7 mg

FeSO4•7H2O
or
Fe2(SO4)3•nH2O

 
2.28 mg
 

KOH

866.7 mg

Milli-Q Water

~100 mL

 

-Dissolve KOH in 31 mL Milli-Q
-Add Na2EDTA•2H2O
-In separate container, dissolve FeSO4•7H2O or Fe2(SO4)3•nH2O in 60.67 mL Milli-Q
-Mix both solutions together
-Bubble filtered air through combined solutions until the color darkens (~2.5 hr)

 

+Pi Stock

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