Steps

Step Solution Medium
For Bacillus strains the addition of 10.0 mg MnSO4 x H2O is recommended for sporulation. 1 465i
Adjust pH to 7.0. For Bacillus strains the addition of 10.0 mg MnSO4 x H2O is recommended for sporulation. 3 1a
Rehydrate and grow lyophilized cells from the ampoule in 5 ml liquid broth 1. Subsequent subculturing may be carried out in liquid medium or on agar plates. 3 1a
Adjust pH to 7.0. 5630 1774
To medium 1 add 20 g/l urea before autoclaving. Do not adjust pH; pH raises to about 8 due to heat degraded urea. 4 2
For sporulation enhancement add 10 mg/l MnSO4 x H2O. 4 2
Adjust pH to 7.3. 5 3
Sterilize glucose and mannitol separately (in 50 ml H2O) and add to the medium after autoclaving. Calcium carbonate in the medium serves as a buffer. The calcium carbonate will settle in agar plates before the agar has set, producing an opaque layer in the bottom. As the strain grows and acid is produced this will react with the calcium carbonate, causing it to dissolve and form zones of clearing immediately below the colonies. 5 3
Adjust pH to 7.0 7 7
Adjust pH to 6.8. 8 8
Sterilize vitamin B12 separately by filtration. Prepare and store yeast cells as autoclaved stock suspension (5 g baker's yeast/100 ml distilled water, adjust pH to 6.5 and autoclave). Adjust pH of medium to 7.2 with KOH before, and after autoclaving and cooling to 50°C (use pH-indicator paper). 9 9
Cultures of myxobacteria delivered freeze-dried: Please see our video tutorial and follow the special instructions: 'Reactivation of Myxobacteria' given with the strain entry of our catalogue. For suspending the freeze-dried cells from ampoules, add about 0.5 - 1.0 ml medium MD1 (per liter: casiton 3.0 g; CaCl2 x 2 H2O, 0.7 g; Mg2SO4 x 7 H2O, 2.0 g) to the vial with freeze dried material. 9 9
Cultures of myxobacteria delivered as active cultures (growing on agar plates): Always use the rim of the swarm as inoculum for fresh media. If the swarms are creamy, transfer high amounts of cell mass to several spots on fresh VY/2 agar medium. If the swarm adheres to the agar or grows within the agar, cut small agar cubes from the rim of the swarm and place onto a fresh agar plate using an appropriate tool such as a lancet. Make sure that the pieces of swarm colonies grown on the agar are transferred to the agar plate. Attempt to place the inoculum in such a way that the swarms are in contact with the fresh agar plate. 9 9
Incubate for up to 3 weeks (in particular Sorangium and Nannocystis strains) at the temperature given for the strain, taking measures against desiccation. If there is no growth after ten days, carefully split up the agar-culture-cubes and squeeze the material to the agar plate and reincubate. 9 9
Sterilize vitamin B12 separately by filtration. Prepare and store yeast cells as autoclaved stock suspension (5 g baker's yeast/100 ml distilled water, adjust pH to 6.5 and autoclave). 10 9a
Adjust pH to 7.3. 5866 545b
The same as Caso Bouillon (Merck or Oxoid, Germany). 5866 545b
Distribute into Hungate tubes and autoclave 257 147
Distribute into Hungate tubes and autoclave. 307 181
Incubate microaerobic, for example in a candle jar. 11 10
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