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Revision as of 11:55, 28 September 2007 (view source) Craig (Talk | contribs) (→Primary & secondary Reagents Required including controls) ← Older edit		Revision as of 11:58, 28 September 2007 (view source) Craig (Talk | contribs) (→Primary & secondary Reagents Required including controls) Newer edit →
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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)

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Revision as of 11:58, 28 September 2007

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• 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 Ampicillin 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

• 50mL Falcon tubes
• Pipette and tips
• 37degree shaker

References