Stock Reagents

Media Recipes

LB Media (for 1L):

• 10 g tryptone peptone
• 5 g yeast extract
• 5 g NaCl
• 1 mL 1N NaOH or 200 uL 5M NaOH
• Add H₂O to 1L
• Autoclave to sterilize
• Store at RT until used

TB Media (for 1L):

• 12 g tryptone peptone
• 24 g yeast extract
• 4 mL glycerol
• Add H₂O to 1L
• Adjust pH to 7.0 with 5M NaOH
• Autoclave to sterilize
• Store at RT until used
• *TB media is used for dense colonies

SOC (for 1L):

• 20 g tryptone peptone
• 5 g yeast extract
• 0.5 g NaCl
• 47 mg KCl
• 0.952 g MgCl₂
• 1.203 g MgSO₄
• 3.6 g glucose
• Autoclave to sterilize
• Store at RT until used

Solutions

10X TAE (for 1L):

• 900 mL dH₂O
• 48.4 g Tris base
• 11.4 mL glacial acetic acid
• 3.72 g EDTA
• H₂O to 1L
• Dilute 1:10 to use 1X TAE

1% agarose:

• 1 g agarose for every 100 mL of 1X TAE
• Microwave until completely clear, boiling
• About 3 min, loosen the cap to allow vapor to escape
• Pour immediately

Culture plates:

• 1.5% (w/v) Bacto agar to LB
• Autoclave to sterilize
• Allow to cool in 50°C water bath until the bottle can be comfortably held
• Add 1uL 1000X antibiotic for every mL of agar made
• Mix gently to avoid making bubbles
• Pour immediately into Petri dishes
• Partially cover with lids to set at RT
• Cover in aluminum foil, store lid-down at 4°C