Team:Alberta/Notebook/protocols/LB

From 2010.igem.org

(Difference between revisions)
(Procedure:)
Line 26: Line 26:
** DON`T put tubes of antibiotics back into freezer.
** DON`T put tubes of antibiotics back into freezer.
* Add the following to 800 ml MilliQ H20:
* Add the following to 800 ml MilliQ H20:
-
{|
+
::Bacto-tryptone || 10g
-
|Bacto-tryptone || 10g
+
::Yeast extract || 5g
-
|-
+
::NaCl || 10g
-
|Yeast extract || 5g
+
 
-
|-
+
-
|NaCl || 10g
+
-
|}
+
* Adjust pH to 7.5 with NaOH (Optional).
* Adjust pH to 7.5 with NaOH (Optional).
* Add 15g agar.
* Add 15g agar.
Line 40: Line 37:
* Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.  
* Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.  
* Pour plates, making sure they are color coded with the type of antibiotic that you used.
* Pour plates, making sure they are color coded with the type of antibiotic that you used.
-
 
==Making LB Broth:==
==Making LB Broth:==

Revision as of 17:57, 26 October 2010

TEAM ALBERTA

Protocol: Making LB Plates

Procedure:

LB Agar (To Make 1L):

  • Before you start:
    • Use AGAR not agarose!
    • Use an Erlenmeyer flask that is twice the volume of your solution (avoid spillage in autoclave).
    • Keep your flask on a stir plate with a stir rod.
    • Put Aluminum foil on the top of the flask and put autoclave tape on.
    • DON`T let it solidify after autoclaving by keeping it on a stir plate while it cools.
    • DON`T put tubes of antibiotics back into freezer.
  • Add the following to 800 ml MilliQ H20:
Bacto-tryptone || 10g
Yeast extract || 5g
NaCl || 10g
  • Adjust pH to 7.5 with NaOH (Optional).
  • Add 15g agar.
  • Melt agar into solution in the microwave (Optional).
  • Adjust volume to 1L with MilliQ H20
  • Autoclave on the liquid cycle (no drying time) (aka LIQ20) for 30 minutes.
  • Wait for it to cool to touchable temperature and then add antibiotics if necessary. Amount of antibiotic required should be in the protocol binder.
  • Pour plates, making sure they are color coded with the type of antibiotic that you used.

Making LB Broth:

  • All of the above WITHOUT AGAR.


Back