Melbourne/Growing up cells
From 2007.igem.org
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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 | + | #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 | + | ##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 | + | #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
- Aliquot 5mL LB into 50ml Falcon tube.
- Add selective antibiotic in amount required for desired concentration.
- Select single colony from Agar Plate and introduce to LB.
- 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
