Team:Newcastle/Growing an overnight cultures

From 2010.igem.org

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(Procedures)
(Procedures)
 
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* LB Broth
* LB Broth
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* Colonies of bacteria
+
* Colonies of bacteria on plate
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* Flame loop
+
* Innoculation loop
* Universal tube
* Universal tube
* Pipette
* Pipette
Line 13: Line 13:
==Procedures==
==Procedures==
-
# 5ml of LB broth is pipetted into the universal tube.
+
# Transfer 5 ml of LB or appropriate media into a universal tube.
-
# The flame loop is used to pick a colony present on the agar plate.
+
# Heat the innoculation loop till red hot to sterilize the loop.
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# The colony is then mixed into the LB Broth in the universal tube.
+
# Allow the innoculation loop to cool down for approximately 30 seconds.  
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# The culture is left overnight at 37°C (''E.coli'', ''B.subtilis'' lower temp)
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# The innoculation loop is then used to pick a single colony of bacteria on the agar plate.
 +
# Innoculate the LB or appropriate media with the bacteria.
 +
# Resterilize the loop.
 +
# The culture is then left overnight at 37°C in a shaking incubator at 200 rpm.

Latest revision as of 14:35, 26 October 2010

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Growing an Overnight Culture

Materials Required

  • LB Broth
  • Colonies of bacteria on plate
  • Innoculation loop
  • Universal tube
  • Pipette

Procedures

  1. Transfer 5 ml of LB or appropriate media into a universal tube.
  2. Heat the innoculation loop till red hot to sterilize the loop.
  3. Allow the innoculation loop to cool down for approximately 30 seconds.
  4. The innoculation loop is then used to pick a single colony of bacteria on the agar plate.
  5. Innoculate the LB or appropriate media with the bacteria.
  6. Resterilize the loop.
  7. The culture is then left overnight at 37°C in a shaking incubator at 200 rpm.


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