Team:Michigan/Media Recipes

From 2010.igem.org

(Difference between revisions)
Line 81: Line 81:
*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
 +
 +
'''40% glucose solution'''
 +
*20 g of glucose
 +
*50 mL of DI water
 +
''filter sterilize''
'''Bushnell-Haas Media with glucose'''
'''Bushnell-Haas Media with glucose'''

Revision as of 20:02, 3 August 2010


Michigan Header




Media Recipes

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

Cyclohexanecarboxylic acid

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

40% glucose solution

  • 20 g of glucose
  • 50 mL of DI water

filter sterilize

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