Team:Groningen/Notebook/5 August 2009
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Revision as of 07:35, 5 August 2009 by Verhoeven1981 (Talk | contribs)
It's my birthday!!! (Michael)
So cake it is....
Wet
GVP Cluster
- → TODO buy a nice cake and eat it ;)
- → TODO isolate plasmids from overnight precultures
- → TODO perform a ligation between cut vector and GVP fragment
- → TODO transform E.coli TOP10 competent cells with ligation product
Ligation
- 2 uL Ligase buffer
- 1 ul T4 Ligase
- 20 uL plasmid pSB1AC3 digested with PstI and SpeI
- 6 uL insert GVP restricted with XbaI and PstI
Incubate:
- 16°C 60min.
- 65°C 10min.
Tranformation
- add 15 uL of the pSB1AC3-GVP ligation product to 50uL competent E.coli TOP10 cells.
Incubate:
- 30 min @ ice
- 5 min 37°C
- 5min @ ice
- add 800uL LB-medium
- incubate for 1 h at 37°
- plate on LB-AMP plates
Dry
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