Dandekar & Chandler: LB

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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....)
(Ingredients)
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
-
#15g agar
+
#15 g/L agar
 +
 
See also: [[Silver: LB Liquid]]
See also: [[Silver: LB Liquid]]

Revision as of 18:01, 27 March 2014

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/L Bacto-tryptone
  2. 5 g/L yeast extract
  3. 5 g/L NaCl

For plates also add

  1. 15 g/L agar

See also: Silver: LB Liquid

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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