LB
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== Summary == | == Summary == | ||
| - | Luria-Bertani Medium (aka L-Broth or LB Medium). | + | 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 == | == Ingredients == | ||
| + | |||
#10 g Bacto-tryptone | #10 g Bacto-tryptone | ||
#5 g yeast extract | #5 g yeast extract | ||
#10 g NaCl | #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== | ==Protocol== | ||
#Mix dry ingredients and add distilled water up to 1 Liter | #Mix dry ingredients and add distilled water up to 1 Liter | ||
| Line 11: | Line 20: | ||
#* 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. | + | 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== | ||
Adapted From: | 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 | J. Sambrook, D.W. Russell, ''Molecular Cloning: A Laboratory Manual'' (Cold Spring Harbor Laboratory Press, New York, ed. 3, 2001) pg. A2.2 | ||
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| + | [[Category:Material]] [[Category:Standard Media]] | ||
Current revision
Contents |
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
