Team:RHIT/Protocols/MediaRecipes

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LB Medium
To 600 ml of DI H₂O, add the following: 6 g tryptone, 3 g yeast extract, 6 g NaCl
Shake until the solutes are dissolved, and adjust the pH to 7.0 with 5 N NaOH. Sterilize by autoclaving on the liquid cycle at 121°C for 35 minutes.

5X M9 Salt solution and M9 Medias
M9 Salt Solution
Dissolve the following into DI H₂O to a final volume of 1L:

• 64g Na₂HPO₄*7 H₂O
• 15 g KH₂PO₄
• 25. g NaCl
• 5.0 g NH₄Cl

Autoclave on liquid cycle to sterilize.

M9 minimal media, M9 minimal + his, M9 minimal + lac, and M9 minimal + lac + his
To 300 ml DI H₂ add the following:

• 61.2 ml M9 salt solution
• .61 ml 1M MgSO₄
• 6.12 ml 20% glucose
• 0.031 ml 1M CaCl₂
• 0.031 g 1% thiamine (vitamin B)
• 0.06 g uracil
• 10 g agar
• .132 g of lac or his (for +his, +lac, and +lac+his only!)

Autoclave at 121°C for 35 minutes.
Prepare MgSO₄, CaCl₂, and glusoce solutions separately and sterilize by autoclaving on the liquid cycle (glucose may be sterilized by passing it through a 0.22 micrometer filter), then add them to the diluted M9 mixture.

SOC Medium
To 600 ml of DI H₂O, add the following:

• 12 g tryptone
• 3 g yeast extract
• 0.3 g NaCl

Shake until the solutes have dissolved, and then add 10 ml of 250 mM solution of KCl. Adjust the pH of the medium to 7.0 with 5 N NaOH. Sterilize by autoclaving on the liquid cycle at 121°C for 35 minutes. After autoclaving, allow the medium to cool to 60°C or less, and add 20 ml of sterile 1M glucose. Just before use, add 5 ml of an autoclaved solution of 2M MgCl₂.