"
Team:Cambridge/Protocols/MediumB
From 2012.igem.org
(Difference between revisions)
(→Medium B) |
|||
| Line 3: | Line 3: | ||
==Medium B== | ==Medium B== | ||
| - | ''10X Medium A base:'' | + | '''10X Medium A base:''' |
*Yeast extract 10g | *Yeast extract 10g | ||
| Line 11: | Line 11: | ||
*50% glucose, filter sterilized 100mL | *50% glucose, filter sterilized 100mL | ||
| - | ''10X Bacillus salts: | + | '''10X Bacillus salts:''' |
*(NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>.3H<sub>2</sub>O 20g | *(NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>.3H<sub>2</sub>O 20g | ||
| Line 20: | Line 20: | ||
*SDW to 1000mL | *SDW to 1000mL | ||
| - | ''Medium A'' | + | '''Medium A''' |
*Sterile water 81mL | *Sterile water 81mL | ||
| Line 27: | Line 27: | ||
*L-Tryptophan (11mg/mL) 0.1mL | *L-Tryptophan (11mg/mL) 0.1mL | ||
| - | ''Medium B'' | + | '''Medium B''' |
*Medium A 10mL | *Medium A 10mL | ||
| Line 33: | Line 33: | ||
*250nM MgCl<sub>2</sub>.6H<sub>2</sub>O 0.1mL | *250nM MgCl<sub>2</sub>.6H<sub>2</sub>O 0.1mL | ||
| - | ''Important'' | + | '''Important''' |
*Autoclave Medium A base before adding glucose, and autoclave Bacillus salts. | *Autoclave Medium A base before adding glucose, and autoclave Bacillus salts. | ||
Revision as of 11:53, 14 September 2012
Medium B
10X Medium A base:
- Yeast extract 10g
- Casamino acids 2g
- Distilled water to 900mL
- Autoclave then add:
- 50% glucose, filter sterilized 100mL
10X Bacillus salts:
- (NH4)2SO4.3H2O 20g
- K2HPO4.3H2O 183g
- KH2PO4 60g
- Tri-sodium citrate 10g
- MgSO4.7H2O 2g
- SDW to 1000mL
Medium A
- Sterile water 81mL
- 10X Medium A base 10mL
- 10X Bacillus salts 9mL
- L-Tryptophan (11mg/mL) 0.1mL
Medium B
- Medium A 10mL
- 50mM CaCl2.2H2O 0.1mL
- 250nM MgCl2.6H2O 0.1mL
Important
- Autoclave Medium A base before adding glucose, and autoclave Bacillus salts.
- Store aliquots of 10X Medium A base 10mL and 10X Bacillus salts 9mL and keep them in the fridge, never use them twice to avoid contamination
