Melbourne/Growing up cells

From 2007.igem.org

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(→Primary & secondary Reagents Required including controls)
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====Method from primary and secondary reagents====
====Method from primary and secondary reagents====
=====Primary & secondary Reagents Required including controls=====
=====Primary & secondary Reagents Required including controls=====
-
*[[Melbourne secondary reagent LB|LB]] (cupboard)
+
*[[Melbourne/Secondary Reagent LB|LB]] (cupboard)
*Selective Antibiotic (minus 20 freezer)
*Selective Antibiotic (minus 20 freezer)
*Plate of transformed cells (cool room)
*Plate of transformed cells (cool room)
=====Method including controls=====
=====Method including controls=====
-
#Aliquot 5mL LB into 50mL falcon tube.
+
#Aliquot 5mL [[Melbourne/Secondary Reagent LB|LB]] into [[Melbourne/primary falcon|50ml Falcon tube]].
#Add selective antibiotic in amount required for desired concentration.
#Add selective antibiotic in amount required for desired concentration.
-
##Add 10uL Ampicillin Stock (5mg/mL) leads to final concentration of 100ug/mL.
+
##Add 10uL [[Melbourne/primary AMP|Ampicilin]] Stock (5mg/mL) leads to final concentration of 100ug/mL.
-
##Add 5uL Kanamycin Stock (5mg/mL) leads to final concentration of 50ug/mL.
+
##Add 5uL [[Melbourne/primary Kan|Kanamycin]] Stock (5mg/mL) leads to final concentration of 50ug/mL.
-
#Select single colony from agar plate and introduce to LB.  
+
#Select single colony from [[Melbourne/Secondary Reagent Agar Plates|Agar Plate]] and introduce to [[Melbourne/Secondary Reagent LB|LB]].  
#Incubate at 37degrees with shaking for approximately 18hrs (varies depending on growth rate of cells)
#Incubate at 37degrees with shaking for approximately 18hrs (varies depending on growth rate of cells)
 +
=====Equipement Required=====
=====Equipement Required=====
-
*50mL Falcon tubes
+
*[[Melbourne/primary falcon|50mL Falcon tubes]]
-
*Pipette and tips
+
*Pipette and [[Melbourne/primary tips|Tips]]
*37degree shaker
*37degree shaker
 +
=====References=====
=====References=====
*
*
__NOTOC__
__NOTOC__

Latest revision as of 12:03, 28 September 2007

<Return to list of protocols> <Team home page>

  • Applications:
    • Miniprep
    • Glycerol stocks
  • Time to complete protocol:
    • Lab time: 15min
    • Waiting time:18hours
  • Approximate cost of materials: $

Method from primary and secondary reagents

Primary & secondary Reagents Required including controls
  • LB (cupboard)
  • Selective Antibiotic (minus 20 freezer)
  • Plate of transformed cells (cool room)
Method including controls
  1. Aliquot 5mL LB into 50ml Falcon tube.
  2. Add selective antibiotic in amount required for desired concentration.
    1. Add 10uL Ampicilin Stock (5mg/mL) leads to final concentration of 100ug/mL.
    2. Add 5uL Kanamycin Stock (5mg/mL) leads to final concentration of 50ug/mL.
  3. Select single colony from Agar Plate and introduce to LB.
  4. Incubate at 37degrees with shaking for approximately 18hrs (varies depending on growth rate of cells)
Equipement Required
References


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