Melbourne/Growing up cells

From 2007.igem.org

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[[Melb:Protocols for Standard Methods|<Back to Protocols page>]]
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[[Melb:Protocols for Standard Methods |<Return to list of protocols>]]  [[Melbourne|  <Team home page>]]
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*Applications:
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**Miniprep
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**Glycerol stocks
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#Aliquot 5mL LB into 50mL falcon tube.
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*Time to complete protocol:
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**Lab time: 15min
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**Waiting time:18hours
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*Approximate cost of materials: $
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====Method from primary and secondary reagents====
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=====Primary & secondary Reagents Required including controls=====
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*[[Melbourne/Secondary Reagent LB|LB]] (cupboard)
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*Selective Antibiotic (minus 20 freezer)
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*Plate of transformed cells (cool room)
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=====Method including controls=====
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#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.
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##Add 10uL Ampicillin Stock (5mg/mL) leads to final concentration of 100ug/mL.
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##Add 10uL [[Melbourne/primary AMP|Ampicilin]] Stock (5mg/mL) leads to final concentration of 100ug/mL.
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##Add 5uL Kanamycin Stock (5mg/mL) leads to final concentration of 50ug/mL.
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##Add 5uL [[Melbourne/primary Kan|Kanamycin]] Stock (5mg/mL) leads to final concentration of 50ug/mL.
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#Select single colony from agar plate and introduce to LB.  
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#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)
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=====Equipement Required=====
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*[[Melbourne/primary falcon|50mL Falcon tubes]]
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*Pipette and [[Melbourne/primary tips|Tips]]
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*37degree shaker
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=====References=====
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*
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__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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