Julius B. Lucks/M9 Supplemented Minimal Media: Difference between revisions
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This protocol is a derivative of [[Endy:M9_media/supplemented]]. Endy protocol items in black, our protocol items in red. | This protocol is a derivative of [[Endy:M9_media/supplemented]]. Endy protocol items in black, our protocol items indented and in <font color=red>red</font>. | ||
==Protocol== | ==Protocol== | ||
For 1L of media | For 1L of media | ||
*500ml 2x[[M9 salts]] | *500ml 2x[[M9 salts]] | ||
** used 11.28g of M9 Minimal Salts in 500ml of MiliQ-filtered H<sub>2</sub>O | ** <font color=red>used 11.28g of M9 Minimal Salts in 500ml of MiliQ-filtered H<sub>2</sub>O</font> | ||
*30ml 10 mg/ml thiamine hydrochloride | *30ml 10 mg/ml thiamine hydrochloride | ||
** used 300 mg thiamine hydrochroride | ** <font color=red>used 300 mg thiamine hydrochroride</font> | ||
*10ml 40% glycerol | *10ml 40% glycerol | ||
** used 8ml 50% glycerol | ** <font color=red>used 8ml 50% glycerol</font> | ||
*20ml 10% casamino acids | *20ml 10% casamino acids | ||
** wasn't sure if this was by weight or not. Assumed 10% by weight, so used 2g casamino acids. | ** <font color=red>wasn't sure if this was by weight or not. Assumed 10% by weight, so used 2g casamino acids.</font> | ||
*20ml 0.1M MgSO<sub>4</sub> | *20ml 0.1M MgSO<sub>4</sub> | ||
** used 0.493 g (FW 246.49 g/mol) | ** <font color=red>used 0.493 g (FW 246.49 g/mol)</font> | ||
*200μl 0.5M CaCl<sub>2</sub> | *200μl 0.5M CaCl<sub>2</sub> | ||
** used the same (made 1ml 0.5 M CaCl<sub>2</sub> with 74 mg (FW 147.01 g/mol)) | ** <font color=red>used the same (made 1ml 0.5 M CaCl<sub>2</sub> with 74 mg (FW 147.01 g/mol))</font> | ||
*419.8ml sterile deionized H<sub>2</sub>O | *419.8ml sterile deionized H<sub>2</sub>O | ||
** we made the above solution and filled to 1L with MiliQ-filtered H<sub>2</sub>O | ** <font color=red>we made the above solution and filled to 1L with MiliQ-filtered H<sub>2</sub>O</font> | ||
We combined the above ingredients, and filtered through a milipore steritop filter into a clean auto-claved 1L bottle. Stored at 4°C | We combined the above ingredients, and filtered through a milipore steritop filter into a clean auto-claved 1L bottle. Stored at 4°C | ||
Latest revision as of 16:00, 3 April 2008
This protocol is a derivative of Endy:M9_media/supplemented. Endy protocol items in black, our protocol items indented and in red.
Protocol
For 1L of media
- 500ml 2xM9 salts
- used 11.28g of M9 Minimal Salts in 500ml of MiliQ-filtered H2O
- 30ml 10 mg/ml thiamine hydrochloride
- used 300 mg thiamine hydrochroride
- 10ml 40% glycerol
- used 8ml 50% glycerol
- 20ml 10% casamino acids
- wasn't sure if this was by weight or not. Assumed 10% by weight, so used 2g casamino acids.
- 20ml 0.1M MgSO4
- used 0.493 g (FW 246.49 g/mol)
- 200μl 0.5M CaCl2
- used the same (made 1ml 0.5 M CaCl2 with 74 mg (FW 147.01 g/mol))
- 419.8ml sterile deionized H2O
- we made the above solution and filled to 1L with MiliQ-filtered H2O
We combined the above ingredients, and filtered through a milipore steritop filter into a clean auto-claved 1L bottle. Stored at 4°C
