Transformation of Bacillus subtilis

Materials required

1. Bacillus subtilis 168
2. 10 µl of DNA
3. SMM medium (1 liter)
   • 2.0 g of ammonium sulphate
   • 14.0g of dipotassium hydrogen phosphate
   • 6.0g of potassium dihydrogen phosphate
   • 1.0g of sodium citrate dehydrate
   • 0.2g of magnesium sulphate
   • Top up the rest of the medium with water
4. MM competence medium
   • 10 ml of SMM medium
   • 125 µl of Solution E (40% glucose)
   • 100 µl of Tryptophane solution (at a concentration of 2mg/ml) –
   • 60 µl of Solution F (1M MgSO4)
   • 10 µl of 20% Casamino acids
   • 5 µl of 0.22% Fe-NH4-citrate
5. Starvation medium
   • 10 ml of SMM medium
   • 125 µl of Solution E (40% glucose)
   • 60 µl of Solution F (1M MgSO4)

Protocol

This protocol will stretch for 2 days.

Protocol for Day 1

1. Inoculate a single colony of Bacillus subtilis 168 into a 15 ml falcon tube containing 10 ml of MM competence media.
2. For control transfer 10 ml of MM competence media without the bacteria.
3. Incubate overnight in a shaking incubator at 37ºC.

Protocol for Day 2

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