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Team:Brown/Notebook/June21
From 2010.igem.org
Monday, June 21 2010
Made LB agar; followed protocol found [http://www.eeb.ucla.edu/Faculty/Barber/Web%20Protocols/LB Agar Plates.pdf]
Autoclaved after boiling solution (until it looked clear).
Ampicillin Stock Solution (James)
- 0.08 g ampicillin trihydrate
- 50 mL dH2O
- 50 mL EtOH
800 µg/mL
Placed stock solution at -20°C
IPTG Stock Solution (also store in fridge)
Aliquots should be stored at -20°C and thawed only 3 to 4 times.
1M stock solution:
- Dissolve 2.38 g IPTG in 8 mL dH2O
- Bring to final volume of 10 mL with dH2O
- Filter sterilize (is this necessary?)
- Store in 1mL aliquots at -20°C.
0.1 mM IPTG corresponds to 0.0119 g IPTG per 0.5 liter
Every 1 mL has 0.238 g (1 M stock)
0.238 g / 0.0119 g = 20
Chloramphenicol Stock Solution
- 2.7216 g of chloramphenicol solid
- 80 mL EtOH
- Store in 1 mL aliquots at -20°C.
Motility Stab LB Agar Recipe
- 500 mL dH2O
- 1.34 g tryptone
- 0.67 g yeast extract
- 1.34 g NaCl
- 2.00 g agar
- Bring into solution; boil for up to 1 minute.
- Autoclave
- Add dissolved media into a culture tube.
Buffer for CaCl2 competence protocols
TfBI – 500 mL total
- 30 mM KoAc = 1.472 g
- 50 mM MnCl2*4H2O = 4.948 g
- 100 mM KCl = 3.728 g
- 10 mM CaCl2*2H2O = 0.735 g
- 15% glycerol = 75 mL
- dH2O = 425 mL
TfBII – 50 mL
- 10 mM Na-MOPS (1M stock) = 0.5 mL
- 75 mM CaCl2*2H2O = 0.5513 g
- 10 mM KCl = 0.0373 g
- 15% glycerol = 7.5 mL
- dH2O = 42.5 mL
L. lactis
6:40 PM – L. lactis put into 30°C incubator
