Melbourne/Transformation Protocol
From 2007.igem.org
(Difference between revisions)
| Line 1: | Line 1: | ||
| - | [[Melb:Protocols for Standard Methods|< | + | [[Melb:Protocols for Standard Methods |<Return to list of protocols>]] [[Melbourne| <Team home page>]] |
| + | *Applications: | ||
| + | *#Amplification of Biobrick DNA for storage and use. | ||
| + | *#Selection aand amplification of ligated constructs | ||
| + | *Time to complete protocol: | ||
| + | **Lab time: 10min, 10min, 10min, 15min. | ||
| + | **Waiting time: 45min, 15min, 1hour, overnight. | ||
| + | *Approximate cost of materials: $ | ||
| + | |||
| + | ====Method from primary and secondary reagents==== | ||
| + | =====Primary & secondary Reagents Required including controls===== | ||
| + | *Competent cells | ||
| + | *DNA for transformation | ||
| + | *LB | ||
| + | *LB-agar plates with selective antibiotic | ||
| + | |||
| + | =====Method including controls===== | ||
#Add 1uL resuspended plasmid DNA to 50uL competent cells. | #Add 1uL resuspended plasmid DNA to 50uL competent cells. | ||
#Incubate on ice for 45min. | #Incubate on ice for 45min. | ||
| Line 12: | Line 28: | ||
#Incubate plate overnight at 37 degrees. | #Incubate plate overnight at 37 degrees. | ||
#Place in cold room until needed. | #Place in cold room until needed. | ||
| + | =====Equipement Required===== | ||
| + | *1.5mL Microfuge tubes | ||
| + | *Ice box | ||
| + | *Pipettes | ||
| + | *42 degree water bath | ||
| + | *37 degree incubator | ||
| + | *Bunsen burner | ||
| + | *Spreader | ||
| + | =====References===== | ||
| + | * | ||
| + | |||
| + | |||
| + | *'''[[User:PhillipDodson|PhillipDodson]] 03:48, 1 July 2007 (EDT)''':Secondary Reagent Template 1/July/2007 | ||
| + | |||
| + | __NOTOC__[[Melb:Protocols for Standard Methods|<Back to Protocols page>]] | ||
Revision as of 05:26, 2 July 2007
<Return to list of protocols> <Team home page>
- Applications:
- Amplification of Biobrick DNA for storage and use.
- Selection aand amplification of ligated constructs
- Time to complete protocol:
- Lab time: 10min, 10min, 10min, 15min.
- Waiting time: 45min, 15min, 1hour, overnight.
- Approximate cost of materials: $
Method from primary and secondary reagents
Primary & secondary Reagents Required including controls
- Competent cells
- DNA for transformation
- LB
- LB-agar plates with selective antibiotic
Method including controls
- Add 1uL resuspended plasmid DNA to 50uL competent cells.
- Incubate on ice for 45min.
- Heat shock in water bath at 42 degrees for 1min.
- Incubate on ice for 15min.
- Add 1mL LB.
- Incubate at 37degrees for 1 hour.
- Spin down cells and remove majority of LB.
- Resuspend cells in remaining LB.
- Under a bunsen spread resuspended bacteria on agar plate on selective antibiotic.
- Incubate plate overnight at 37 degrees.
- Place in cold room until needed.
Equipement Required
- 1.5mL Microfuge tubes
- Ice box
- Pipettes
- 42 degree water bath
- 37 degree incubator
- Bunsen burner
- Spreader
References
- PhillipDodson 03:48, 1 July 2007 (EDT):Secondary Reagent Template 1/July/2007
