LB: Difference between revisions
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#10 g NaCl | #10 g NaCl | ||
For plates also add | |||
#15g agar | |||
See also: [[Silver: LB Liquid]] | See also: [[Silver: LB Liquid]] | ||
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#* 250°'''F''', 22psi, 30 minutes | #* 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). | 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== | ==Source== | ||
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J. Sambrook, D.W. Russell, ''Molecular Cloning: A Laboratory Manual'' (Cold Spring Harbor Laboratory Press, New York, ed. 3, 2001) pg. A2.2 | J. Sambrook, D.W. Russell, ''Molecular Cloning: A Laboratory Manual'' (Cold Spring Harbor Laboratory Press, New York, ed. 3, 2001) pg. A2.2 | ||
[[Category:Material]] [[Category:Standard Media]] |
Revision as of 14:23, 23 December 2011
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 Bacto-tryptone
- 5 g yeast extract
- 10 g NaCl
For plates also add
- 15g agar
See also: Silver: LB Liquid
Note: There are two formulations of LB, Miller and Lennox, that differ in the amount of NaCl. Lennox has less salt, only 5 g/L. The Qiagen miniprep kit recommends LB with 10 g NaCl for highest plasmid yields.
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