Smolke Endy LB

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(LB preparation protocol)
(LB preparation protocol)
Line 7: Line 7:
Measure in Bacto-peptone, Yeast, NaCl, and NaOH.
Measure in Bacto-peptone, Yeast, NaCl, and NaOH.
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Add 1900 mls water from the Millipore in the wash room.
+
Add 1900 mls water from the Millipore in the lab.
Mix on the stir plate.
Mix on the stir plate.

Revision as of 14:49, 16 May 2012

LB preparation protocol

Day before, ask how much is needed to make. Log the batch number in your notebook.

Make two liters at a time, in a 2 liter flask, add a stir bar.

Measure in Bacto-peptone, Yeast, NaCl, and NaOH. Add 1900 mls water from the Millipore in the lab.

Mix on the stir plate. Pour into a 2L cylinder and bring up volume to 2L.

Return to flask, and remix.

With funnel, aliquot 100ml into 10x 250ml bottles and 200 ml into 5x 250 ml bottles. Label LB, Lot number and your initials.

Loosely cap, add autoclave tape, and place in autoclave tray with 1-2in of water.

Autoclave on Liquid 30 or Liquid 60 (if making LB agar at the same time)

With orange gloves, INDIVIDUALLY remove each bottle from the water and let cool. When cool, tighten cap and store in chemical room cabinet.

Recipe

For 2 Liters

  • 20 G Bacto-Peptone
  • 10 G Yeast Extract
  • 10 G NaCl
  • 2 ML 1N NaOH
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