TITLE: Recipe for standard BG-11 media
AUTHORS: Anna Behle

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Stanier RY, Deruelles J, Rippka R, Herdman M, Waterbury JB: </span><span style = "font-weight:bold;">Generic Assignments, Strain Histories and Properties of Pure Cultures of Cyanobacteria.</span><span> Microbiology 1979, 111:1–61.</span></div><div class = "text-block"><span>Recipes for standard and alternative BG11 for culturing freshwater cyanobacteria, such as </span><span style = "font-style:italic;">Synechocystis</span><span> sp. PCC 6803, as described.</span></div><div class = "text-block">Media is usually not suitable for marine cyanobacteria.</div><div class = "text-block"> Final Concentration of Medium.</div><div class = "text-block">CaCl2*2 H2O                        0.036 g/L</div><div class = "text-block">Citric acid                              0.006 g/L</div><div class = "text-block">NaNO3                                   1.4958 g/L</div><div class = "text-block">MgSO4* 7 H2O                     0.0749 g/L</div><div class = "text-block">0.25M Na2EDTA (pH 8)      0.0056 mL/L</div><div class = "text-block">Na2CO3                                 20 µg/L</div><div class = "text-block">Fe(III) Ammonium citrate   6 µg/L</div><div class = "text-block">K2HPO4 * 3H2O                   30 µg/L</div><div class = "text-block">TES Buffer (pH 8)                10 mM</div><div class = "text-block">H3BO3                                   2.86 mg/L</div><div class = "text-block">MnCl2 * 4 H2O                     1.81 mg/L</div><div class = "text-block">ZnSO4 * 7 H2O                      0.222 mg/L</div><div class = "text-block">Na2MoO4 * 2 H2O                0.390 mg/L</div><div class = "text-block">Co(NO3)2 *6 H2O                0.049 mg/L</div><div class = "text-block">(CuSO4 * 5 H2O                    0.079 mg/L if required)    </div></div>

[STEPS]
?. [100 x BG11 stock:]
CaCl2 2H2O (3.6 g · L-1)Citric acid (0.6 g · L-1)NaNO3 (149.58 g · L-1)MgSO4 · 7 H2O (7.49 g · L-1)0.25 M Na2-EDTA, pH 8.0  (0.56 ml · L-1) For 100x BG11 Stock -N: Omit NaNO3.For 100x BG11 Stock -N:
?. [Supplemental stocks for standard media:]
1000x Na2CO3: 20 mg mL-1100x TES-buffer, pH 8.0 (1M), adjust with KOH1000x K2HPO4 x 3 H2O: 30 mg · mL-11000x Fe(III) ammonium citrate (6 mg ·m L-1)5000x CuSO4 5 H2O (395 ng · mL-1) (sterilize using a filter)
?. [Trace metal mix:]
1000x concentration:H3BO3 (2.86 g · L-1)MnCl2 · 4 H2O (1.81 g · L-1)ZnSO4 · 7 H2O (0.222 g · L-1)Na2MoO4 · 2 H2O (0.390 g · L-1)Co(NO3)2 · 6 H2O (0.049 g · L-1)For BG11 lacking certain metals (e.g. for working with metal inducible promoters PpetE, PcoaT, PziaA etc., trace metal mix can be prepared lacking these chemicals and used instead of standard trace metal mix.
?. [Standard 1x BG11]
Fill 1 L bottle with 500 mL ultra pure water. Add stock solutions as shown below.  AB1Stock solutionVolume2100x BG11 Stock10 mL31000x Na2CO31 mL41000x K2HPO4 x 3 H2O1 mL5100x TES-buffer10 mL61000x Trace Metal Mix1 mLAdd ultra pure water to 1 L.Autoclave.After autoclaving, add 1 mL 1000x Fe(III) ammonium citrate.Optional: After autoclaving, add 200 µL 5000x CuSO4
AB1Stock solutionVolume2100x BG11 Stock10 mL31000x Na2CO31 mL41000x K2HPO4 x 3 H2O1 mL5100x TES-buffer10 mL61000x Trace Metal Mix1 mL
?. [Standard 1x BG11 -N]
Fill 1 L bottle with 500 mL ultra pure water. Add stock solutions as shown below.  AB1Stock solutionVolume2100x BG11 Stock -N10 mL31000x Na2CO31 mL41000x K2HPO4&nbsp;x 3 H2O1 mL5100x TES-buffer10 mL61000x Trace Metal Mix1 mLAdd ultra pure water to 1 L.Autoclave.After autoclaving, add 1 mL sterile 1000x Fe(III) ammonium citrate.Optional: After autoclaving, add 200 µL sterile 5000x CuSO4
AB1Stock solutionVolume2100x BG11 Stock -N10 mL31000x Na2CO31 mL41000x K2HPO4&nbsp;x 3 H2O1 mL5100x TES-buffer10 mL61000x Trace Metal Mix1 mL
?. [Standard 2x BG11 for agar plates]
Fill 500 mL bottle with 250 mL ultra pure water. Add stock solutions as shown below.  AB1Stock solutionVolume2100x BG11 Stock -N10 mL31000x Na2CO31 mL41000x K2HPO4&nbsp;x 3 H2O1 mL5100x TES-buffer, pH = 8.010 mL61000x Trace Metal Mix1 mLAdd ultra pure water to 500 mL.Autoclave.After autoclaving, add 1 mL sterile 1000x Fe(III) ammonium citrate.Optional: After autoclaving, add 200 µL sterile 5000x CuSO4
AB1Stock solutionVolume2100x BG11 Stock -N10 mL31000x Na2CO31 mL41000x K2HPO4&nbsp;x 3 H2O1 mL5100x TES-buffer, pH = 8.010 mL61000x Trace Metal Mix1 mL
?. [BG11 plates]
Prepare 1.5 % agar: Weigh 4.5 g Bacto Agar. Fill up to 300 mL. Autoclave.Microwave agar until liquid. Let cool.
?. In a 50 mL Falcon, add 1 vol 2x BG11 and 1 vol liquid 1.5 % agar. (Note: Usually, one plate requires 30-40 mL total volume.)
?. When mixture is hand warm, add appropriate antibiotics, if required. Quickly pour plate, avoiding air bubbles.