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Team:TU Darmstadt/Materials/LB
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===Protocol=== | ===Protocol=== | ||
| - | + | ''' For medium ''' | |
#Mix dry ingredients and add dH<sub>2</sub>Oup to 999 mL | #Mix dry ingredients and add dH<sub>2</sub>Oup to 999 mL | ||
#Adjust the pH to 7.0 add the missing amount of water to 1L (5M NaOH works best with small voluminas) | #Adjust the pH to 7.0 add the missing amount of water to 1L (5M NaOH works best with small voluminas) | ||
| Line 58: | Line 55: | ||
#Autoclave | #Autoclave | ||
| - | + | ''' For plates ''' | |
#Mix dry ingredients and add dH<sub>2</sub>O up to less than 1 Liter | #Mix dry ingredients and add dH<sub>2</sub>O up to less than 1 Liter | ||
#After dissolved, add agar | #After dissolved, add agar | ||
| Line 68: | Line 65: | ||
#Let solidify about 30 min (cools faster when placed in singels on the bottom of the flow-bench add (description, date, additive, (owner)) | #Let solidify about 30 min (cools faster when placed in singels on the bottom of the flow-bench add (description, date, additive, (owner)) | ||
#store at 4°C | #store at 4°C | ||
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Latest revision as of 16:41, 1 October 2013
LB
Luria-Bertani Medium (aka LB medium) is one of the most common used standard growth medium for a variety of bacteria.
Ingredients
For 1L:
- 10 g tryptone
- 5 g yeast extract
- 10 g NaCl
For plates add:
- 15g agar (according for a volume of 1L)
Protocol
For medium
- Mix dry ingredients and add dH2Oup to 999 mL
- Adjust the pH to 7.0 add the missing amount of water to 1L (5M NaOH works best with small voluminas)
- Pour the mix into a 2 L flask (or greater), before autoclaving, due to boiling retardation
- Autoclave
For plates
- Mix dry ingredients and add dH2O up to less than 1 Liter
- After dissolved, add agar
- Pour into 2 L flask (or greater)
- Autoclave
- Cool media down to handable temperatures (a laser thermometer is a great gadget to determine 'handable' temperatures)
- Add appropriate antibiotic (usually somewhat between 1-10µl of a 50µM stock)
- Pour the solution on plates under sterile conditions (constant flow) try to use all plates after opening of the packing
- Let solidify about 30 min (cools faster when placed in singels on the bottom of the flow-bench add (description, date, additive, (owner))
- store at 4°C
