Minimal Basal Medium (liquid)
Christa Smith
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Abstract
A defined artificial seawater medium for growing bacteria.
Before start
Acid wash all glassware with 10% HCl and rinse well with dH2O.
Attachments
MBM.docx
Steps
Sea Salt Solution
Add 20 g Sigma Sea Salts into a 1 L glass bottle.
Add 699 mL dH2O to sea salts in bottle and mix well to dissolve.
Set prepared sea salt solution aside until ready to autoclave.
FeEDTA Stock
Add 50 mg FeEDTA into a 250 mL glass bottle.
Add 100 mL dH2O to FeEDTA in bottle and mix well to dissolve (stock concentration is 1.36 mM).
Filter-sterilize FeEDTA solution using a 0.2 micron PES membrane filter.
Basal Medium
Add 150 mL of 1 M Tris HCl, pH 7.5, into a 1 L glass bottle.
Add 0.34 g NH4Cl into the same bottle and dissolve.
Add 182.98 mg K2HPO4 into the same bottle and dissolve.
Add 375 mL dH2O to the same bottle and mix well.
Set prepared basal medium aside until ready to autoclave.
Autoclave
Autoclave sea salt solution and basal medium.
Cool autoclaved solutions to room temperature.
Mix
Add 50 mL of FeEDTA stock to the bottle of sea salt solution and mix well.
Add 250 mL basal medium to the bottle of sea salt solution and mix well.
Add 1 mL of vitamin supplement the the bottle of sea salt solution and mix well.
MBM Vitamin Supplement
Finish
Store prepared medium at 4 Celsius.
Carbon source
A carbon source can be added to the finished minimal medium.
