Team:Newcastle/Transformation of B. subtilis
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#* 10 µl of 20% Casamino acids | #* 10 µl of 20% Casamino acids | ||
#* 5 µl of 0.22% Fe-NH4-citrate | #* 5 µl of 0.22% Fe-NH4-citrate | ||
+ | # Starvation medium | ||
+ | #* 10 ml of SMM medium | ||
+ | #* 125 µl of Solution E (40% glucose) | ||
+ | #* 60 µl of Solution F (1M MgSO4) | ||
Revision as of 15:14, 5 August 2010
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Transformation of Bacillus subtilis
Materials required
- Bacillus subtilis 168
- 10 µl of DNA
- 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
- 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
- Starvation medium
- 10 ml of SMM medium
- 125 µl of Solution E (40% glucose)
- 60 µl of Solution F (1M MgSO4)
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