Protocols
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
