LB

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== Summary ==
== Summary ==
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Luria-Bertani Medium (aka L-Broth or LB Medium).  Standard growth medium for a variety of bacteria and conditions.
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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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== Ingredients ==
== Ingredients ==
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#10 g Bacto-tryptone
#10 g Bacto-tryptone
#5 g yeast extract
#5 g yeast extract
#10 g NaCl
#10 g NaCl
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 +
For plates also add
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#15g agar
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See also: [[Silver: LB Liquid]]
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''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.
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==Protocol==
==Protocol==
#Mix dry ingredients and add distilled water up to 1 Liter
#Mix dry ingredients and add distilled water up to 1 Liter
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#* 250°'''F''', 22psi, 30 minutes
#* 250°'''F''', 22psi, 30 minutes
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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. We usually obtain this from the kitchen. 
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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
 +
 
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===For plates===
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#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
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==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

  1. 10 g Bacto-tryptone
  2. 5 g yeast extract
  3. 10 g NaCl

For plates also add

  1. 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

  1. Mix dry ingredients and add distilled water up to 1 Liter
  2. Pour into 2 L flask (or greater)
  3. 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

  1. Mix dry ingredients and add distilled water up to less than 1 Liter
  2. After dissolved, add agar
  3. Pour into 2 L flask (or greater)
  4. Autoclave (liquid cycle)
    • 250°F, 22psi, 30 minutes
  5. Cool media down to 60-55C (uncomfortable but not painful)
  6. Add appropriate antibiotic
  7. Pour the solution on sterile (???) plates
  8. Let solidify about 1 h, wrap, label (name, date, additive)
  9. 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

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