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From 2011.igem.org

Recipes

L Broth

For 1L:

• 10g Tryptone
• 5g Yeast extract
• 5g NaCl

PYE Broth

For 1L:

• 2g Bacto peptone
• 1g Yeast extract
• 1mL 1M MgSO₄⋅7H₂O
• 1mL 0.5M CaCl₂

M2G Minimal Broth

1. For 1L of 20x M2 Salts:
   • 17.4g Na₂HPO₄
   • 10.6g KH₂PO₄
   • 10g NH₄Cl
2. Autoclave this sol'n.
3. To make M2G:
   1. Dilute 20x M2 salts sol'n to 1x and autoclave.
   2. Add the following sterile sol'ns:
      • 1mL 0.5M MgSo₄
      • 10mL 20% Glucose
      • 1mL 10mM FeSO₄, 10mM EDTA (Sigma #F-0518, kept at 4°C)
      • 500μL 1M CaCl₂ (Note: add this last to avoid precipitation)

Agar Plates

Add 15g Bacto-agar to 1L of liquid media before autoclaving.

Antibiotics Kitchen

Antibiotic	Color Code	Stock (mg/mL)	LB Plate Conc (μg/mL)	PYE Plate Conc (μg/mL)
Ampicillin (amp)	Red	50	100	50
Carbenicillin (carb)	Red	50	50	50
Chloramphenicol (chlor)	2 Orange	5 (in EtOH)	30	1
Kanamycin (kan)	blue	50	50	25
Nalidixic acid (nal)	Green	10	20	20
Tetracycline (tet)	Black	2 (in EtOH)	12	–
Oxytetracycline (oxytet)	Black	2 (in EtOH)	−	2
Spectinomycin (spec)	Purple	20	50	100

Note: for liquid cultures in PYE broth, use half the concentration that would be used for a PYE plate (LB remains the same for liquid and sold cultures).

Freezer Stocks

To an appropriate tube add:

• 700μL overnight liquid culture
• 300μL 50% glycerol sol'n

Then place in storage at -80°C.