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Team:Michigan/Media Recipes
From 2010.igem.org
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Media Recipes | Media Recipes | ||
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*1049 uL of DI water | *1049 uL of DI water | ||
''filter sterilize with .22 um filter'' | ''filter sterilize with .22 um filter'' | ||
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| + | '''40% glucose solution''' | ||
| + | *20 g of glucose | ||
| + | *50 mL of DI water | ||
| + | ''filter sterilize'' | ||
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| + | '''0.1 M NaOH solution ''' | ||
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| + | BE VERY CAREFUL WHEN MIXING! SOLID NaOH BURNS YOUR SKIN BUT YOU MAY NOT FEEL IT AT FIRST! MAKE SURE ANY SPILLS ARE CLEANED UP VERY WELL! | ||
| + | *0.2 g of NaOH | ||
| + | *50 mL of sterile water | ||
| + | ''Filter Sterilize'' | ||
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==Oil Sands Recipes== | ==Oil Sands Recipes== | ||
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*665 uL of 1 M MgSO4 solution | *665 uL of 1 M MgSO4 solution | ||
*72 uL of 1 M CaCl2 solution | *72 uL of 1 M CaCl2 solution | ||
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'''Bushnell-Haas Media with glucose''' | '''Bushnell-Haas Media with glucose''' | ||
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*2 mL of sterile DI water | *2 mL of sterile DI water | ||
*6.25 mL of casamino acid solution | *6.25 mL of casamino acid solution | ||
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'''Bushnell-Haas Media with cyclohexanecarboxylic acid adjusted to pH of 9''' | '''Bushnell-Haas Media with cyclohexanecarboxylic acid adjusted to pH of 9''' | ||
Revision as of 15:18, 4 August 2010
Media Recipes
General Recipies
LB media
- 10 g LB broth
- 500 mL DI water
autoclave to sterilize
LB media with agar
- 10 g LB broth
- 7.5 g agar
- 500 mL DI water
autoclave to sterilize
add antibiotics after autoclaving when warm to touch (about 50°C)
pour plates when warm to the touch (about 50°C)
0.1 M CaCl2 solution
- 0.735 g of CaCl2*2 H20
- 50 mL of DI water
filter sterilize
1.0 M CaCl2 solution
- 7.35 g of CaCl2*2 H20
- 50 mL of DI water
filter sterilize
1.0 M MgSO4
- 12.323 g of MgSO4*7 H20
- 50 mL of DI water
filter sterilize
Ampicillin Antibiotic Stock Solution (100 mg/mL)
- 1 g of ampicillin
- 5 mL of DI water
- 5 mL of ethanol
filter sterilize and make 1 mL aliquotes to store in the -20°C freezer
Kanamycin Antibiotic Stock Solution (50 mg/mL)
- 250 mg of kanamycin
- 5 mL of DI water
filter sterilize and make 1 mL aliquots to store in the -20°C freezer
Tetracycline Antibiotic Stock Solution (50 mg/mL)
- 250 mg of tetracycline
- 2.5 mL of DI water
- 2.5 mL of ethanol
filter sterilize and make 1 mL aliquots to store in the -20°C freezer
IPTG stock solution (1 M)
- 0.25 g of IPTG
- 1049 uL of DI water
filter sterilize with .22 um filter
40% glucose solution
- 20 g of glucose
- 50 mL of DI water
filter sterilize
0.1 M NaOH solution
BE VERY CAREFUL WHEN MIXING! SOLID NaOH BURNS YOUR SKIN BUT YOU MAY NOT FEEL IT AT FIRST! MAKE SURE ANY SPILLS ARE CLEANED UP VERY WELL!
- 0.2 g of NaOH
- 50 mL of sterile water
Filter Sterilize
Oil Sands Recipes
1.5 mg/mL Cyclohexanecarboxylic acid solution
Cyclohexanecarboxylic acid is volatile should only be worked with in a hood. For accurate measurement use covered scale with sliding doors in the hood. Stored in the corrosive cabinet
- 0.075 g cyclohexanecarboxylic acid
- 50 mL of DI water
filter sterilize
Bushnell-Haas Salts (w/out carbon source)
- 0.4 g of KH2PO4
- 0.4 g of K2HPO4
- 0.4 g of (NH4)2 SO4 or NH4NO3
- 0.002 g of FeCl3
- 318 mL of DI H2O
Autoclave at 15 psi and 121C for 30 minutes
After the media has cooled completely add:
- 665 uL of 1 M MgSO4 solution
- 72 uL of 1 M CaCl2 solution
Bushnell-Haas Media with glucose
- 39.75 mL of Bushnell-Haas Salts
- 2 mL of 40% glucose solution
- 8.25 mL of sterile DI water
Bushnell-Haas Media with cyclohexanecarboxylic acid
- 39.75 mL of Bushnell-Haas Salts
- 4 mL of cyclohexanecarboxylic acid solution
- 6.25 mL of sterile DI water
20 mg/mL Casamino Acid Solution
- 0.18 g Casamino acid
- 9 mL of DI water
Filter sterilize
Bushnell-Haas Media with glucose and casamino acids
- 39.75 mL of Bushnell-Haas Salts
- 2 mL of 40% glucose solution
- 2 mL of sterile DI water
- 6.25 mL of casamino acid solution
Bushnell-Haas Media with cyclohexanecarboxylic acid adjusted to pH of 9
- 39.75 mL of Bushnell-Haas Salts
- 4 mL of cyclohexanecarboxylic acid solution
- 6.25 mL of sterile DI water
Test how much 0.1 M NaOH must be added to 1 mL of media to raise it to a pH of 9 (add 1 uL of base at a time)
Scale up the amount of 0.1 M NaOH solution to add to the remaining volume of media (approaching final amount slowly)
On 8/3/2010 approximately 20 uL of 0.1 M NaOH had to be added to 1 mL to get to a pH between 8 and 9 (tested with pH paper) and 675 uL of 0.1 M NaOH was added into the 45 mL of remaining media.
