Dandekar & Chandler: LB: Difference between revisions
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(New page: == Summary == Luria-Bertani Medium (aka L-Broth or LB Medium). (Bertani says LB really stands for lysogeny broth.) LB is a standard growth medium for a variety of bacteria and conditions....) |
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Line 4: | Line 4: | ||
== Ingredients == | == Ingredients == | ||
#10 g Bacto-tryptone | #10 g/L Bacto-tryptone | ||
#5 g yeast extract | #5 g/L yeast extract | ||
#5 g NaCl | #5 g/L NaCl | ||
For plates also add | For plates also add | ||
# | #15 g/L agar | ||
See also: [[Silver: LB Liquid]] | See also: [[Silver: LB Liquid]] | ||
Revision as of 15:01, 27 March 2014
Summary
Luria-Bertani Medium (aka L-Broth or LB Medium). (Bertani says LB really stands for lysogeny broth.) LB is a standard growth medium for a variety of bacteria and conditions.
Ingredients
- 10 g/L Bacto-tryptone
- 5 g/L yeast extract
- 5 g/L NaCl
For plates also add
- 15 g/L agar
See also: Silver: LB Liquid
Protocol
- Mix dry ingredients and add distilled water up to 1 Liter
- Pour into 2 L flask (or greater)
- Autoclave (liquid cycle)
- 250°F, 22psi, 30 minutes
Notes: We do not pH medium when we make it on the fly. However, if it is really important, pH the medium to 7.0 with 5M NaOH (~200µL). Check with pH paper
For plates
- Mix dry ingredients and add distilled water up to less than 1 Liter
- After dissolved, add agar
- Pour into 2 L flask (or greater)
- Autoclave (liquid cycle)
- 250°F, 22psi, 30 minutes
- Cool media down to 60-55C (uncomfortable but not painful)
- Add appropriate antibiotic
- Pour the solution on sterile (???) plates
- Let solidify about 1 h, wrap, label (name, date, additive)
- keep at 4°C
Source
Adapted From:
J. Sambrook, D.W. Russell, Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory Press, New York, ed. 3, 2001) pg. A2.2