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

pH: 8.3

 

 

PES Stock Solution

NaNO3

50 mg

Na2-Glycerolphosphate•4H2O

50 mg

Tris Buffer

500 mg

Iron Solution

25 mL

Trace Metal

15 mL

Milli-Q Water

100 mL

 

-Measure out Milli-Q
-Add NaNO3 and Na2-Glycerolphosphate•4H2O
-Combine trace metal and iron solutions in separate container
-Autoclave both containers
-Once cool, combine both mixtures
-Store at 4oC in 10 mL aliquots

 

Iron Solution

FeNH4(SO4)2•??H2O

???

Na2EDTA

300 mg

Milli-Q Water

~500 mL

 

-Measure out 250 mL Milli-Q
-Add components in order
-Bring volume up to 500 mL
-Autoclave for 25 minutes

 

Trace Metal Solution A5+Co

H3BO3

2.86 g

MnCl2•4H2O

1.81 g

ZnSO4•7H2O

222 mg

Na2MoO4•2H2O

390 mg

CuSO4•5H20

79 mg

Co(NO3)2•6H2O

49 mg

Milli-Q Water

~1 L

 

-Measure out 500 mL Milli-Q
-Add components in order
-Bring volume to 1 L with Milli-Q

 

Vitamin Solution

Biotin

10 mg

Thiamine

200 mg

Cyanocobalamin

10 mg

Folic Acid

10 mg

Myo-Inositol

10 mg

Calcium Pantothenate

100 mg

Milli-Q Water

~1 L

 

-Measure out 500 mL Milli-Q
-Add components in order
-Bring volume up to 1 L with Milli-Q
-Filter sterilize before use