Team:Wash U/Protocol

• 10g Tryptone
• 5g Yeast Extract
• 10g NaCl
• 15g Agar
• up to 1 Liter H20

ATCC medium: Van Niel's yeast agar

• 1.0 g K2HPO4
• 0.5 g MgSO4
• 10.0 g yeast extract
• 20.0 g agar
• 1.0 L tap water
• Adjust pH to 7.0-7.2

Super Optimal Broth (SOB)

• 2% w/v bacto-tryptone (20g)
• 0.5% w/v bacto-yeast extract (5g)
• 8.56mM NaCl (0.5g) or 10mM NaCl (0.584g)
• 2.5mM KCl (0.186g)
• ddH2O to 1L

Super Optimal Broth with Catabolite repression (SOC)

• In addition to all of the SOB components,
• 10mM MgCl2 (0.952g)
• 20mM glucose (3.603g)

TAE Buffer

• 242 g Tris base (2-amino-2-hydroxymethyl-propane-1,3-diol) (= 2 mole)
• 57.1 ml glacial acetic acid (= 100% acetic acid) (57.19 ml = 1 mole)
• 100 ml 0.5 M Na2 EDTA (pH 8.0)
• ddH2O to 1L

TBE Buffer

• 53 g of Tris base (CAS# 37186)
• 27.5 g of boric acid (CAS# 11280)
• 20 ml of 0.5 M EDTA (CAS# 60004) (pH 8.0)

Tris EDTA (TE) buffer

• 750 mL d-H20
• 242 g Tris base
• 57.1 mL glacial acetic acid
• 100 mL 0.5M EDTA (93.05 g EDTA in 500 mL d-H20, pH ~8.0)
• fill to 1 L
• adjust pH to 8.5

Gel (for Electrophoresis)

• 50 mL 1X TAE buffer
• 0.8 g agarose
• 2.5 microliters EtBr
• Note: Jacob suggested adding 1.0 microliter EtBr to gel and 3.0 microliters to TAE buffer in rig.