TITLE: MMH1 Solution
AUTHORS: Erçağ Pinçe

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Minimal medium H1 protocol for bacterial competition experiments</div></div>

[STEPS]
?. [Prepare stock solutions of the salts]
Prepare Potassium Phosphate:   ABCDEFGHI1Name of the chemicalFormulaBrandMolecular Weight(gr/mol)Final ConcentrationStockFactor&nbsp;&nbsp;Stock in 100mL in gr&nbsp;2Potassium phosphate(Monobasic)&nbsp;KH2PO4Sigma-Aldrich139.06&nbsp;50 mM&nbsp;1M&nbsp;20x9.5&nbsp;3Potassium Phosphate(Dibasic)&nbsp;K2HPO4Sigma-Aldrich&nbsp;174.1850 mM&nbsp;1M20x&nbsp;5.25&nbsp;Dissolve the reagents in 80 mL of ddH2OAdjust pH to 7.0Make the volume up to 100 mL with ddH2OFilter sterilize and store at room temperature.  Prepare stock solutions of other salts:   Mix each compound in 200 mL in separate beakers and stir for ~20mins at room temperature.Filter sterilize each solution and store at room temperature. Important Note: You can either use the stock solution of NaCl [1.34M] or you can directly add it to the final MMH1 mix in its solid form. Prepare stock solutions of other salts:  ABCDEFGH1Name of the chemicalFormulaBrandMolecular Weight(gr/mol)Final ConcentrationStock Con.Factor&nbsp;Stock in 200 mL in gr2Ammonium Sulfate(NH4)2SO4Sigma-Aldrich132.147.6mM&nbsp;760mM100x20.093Magnesium Sulfate heptahydrateMgSO4Sigma-Aldrich246.470.5mM&nbsp;50mM100x2.464Sodium ChlorideNaClSigma-Aldrich&nbsp;58.4467mM&nbsp;1.34M&nbsp;20x15.665&nbsp;Iron Sulfate
(Cover with foil)
&nbsp;Fe2(SO4)3&nbsp;Sigma-Aldrich&nbsp;399.881.25µM&nbsp;1.25mM&nbsp;1000x0.1&nbsp;Mix each compound in 200 mL in separate beakers and stir for ~20mins at room temperature.Filter sterilize each solution and store at room temperature. Important Note: You can either use the stock solution of NaCl [1.34M] or you can directly add it to the final MMH1 mix in its solid form.
ABCDEFGHI1Name of the chemicalFormulaBrandMolecular Weight(gr/mol)Final ConcentrationStockFactor&nbsp;&nbsp;Stock in 100mL in gr&nbsp;2Potassium phosphate(Monobasic)&nbsp;KH2PO4Sigma-Aldrich139.06&nbsp;50 mM&nbsp;1M&nbsp;20x9.5&nbsp;3Potassium Phosphate(Dibasic)&nbsp;K2HPO4Sigma-Aldrich&nbsp;174.1850 mM&nbsp;1M20x&nbsp;5.25&nbsp;
ABCDEFGH1Name of the chemicalFormulaBrandMolecular Weight(gr/mol)Final ConcentrationStock Con.Factor&nbsp;Stock in 200 mL in gr2Ammonium Sulfate(NH4)2SO4Sigma-Aldrich132.147.6mM&nbsp;760mM100x20.093Magnesium Sulfate heptahydrateMgSO4Sigma-Aldrich246.470.5mM&nbsp;50mM100x2.464Sodium ChlorideNaClSigma-Aldrich&nbsp;58.4467mM&nbsp;1.34M&nbsp;20x15.665&nbsp;Iron Sulfate
(Cover with foil)
&nbsp;Fe2(SO4)3&nbsp;Sigma-Aldrich&nbsp;399.881.25µM&nbsp;1.25mM&nbsp;1000x0.1&nbsp;
The protocol for preparing 1M potassium phosphate was taken from Cold Spring Harbor Protocolshttp://cshprotocols.cshlp.org/content/2017/7/pdb.rec090290.full?text_only=true
?. [Prepare MMH1 mix]
MMH1: 1) Add the salts and Thiamine435 mL ddH2O5 mL Ammonium Sulfate [100x]5 mL Magnesium Sulfate [100x] 5 mL Iron(III) Sulfate [100x] 25 mL Sodium Chloride [20x] 25 mL Potassium Phosphate [20x] 50 mg Thiamine Hydrochloride (Sigma-Aldrich)  MMH1[2x]: 370 mL ddH2O10 mL Ammonium Sulfate [100x]10 mL Magnesium Sulfate [100x] 10 mL  Iron(III) Sulfate [100x] 50 mL Sodium Chloride [20x]50 mL Potassium Phosphate [20x]100 mg Thiamine Hydrochloride (Sigma-Aldrich)MMH1[2x]: 370 mL ddH2O10 mL Ammonium Sulfate [100x]10 mL Magnesium Sulfate [100x] 10 mL  Iron(III) Sulfate [100x] 50 mL Sodium Chloride [20x]50 mL Potassium Phosphate [20x]100 mg Thiamine Hydrochloride (Sigma-Aldrich)2-) Stir mixes at room temperature for ~30mins. 3-) Measure pH and adjust it to 7.0 if necessary. 4-) Filter sterilize and store in the cold room(+4C)