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Team:Groningen/Notebook/6 July 2009
From 2009.igem.org
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(New page: {{Team:Groningen/Notebook/Day/Header}} '''Transformation of ''E. coli TOP10'' with BBa_J23109, BBa_J23100 and BBa_J23106''' * BBa_J23109 iGEM plate 2, 2G * BBa_J23100 iGEM plate 1, 18C *...) |
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{{Team:Groningen/Notebook/Day/Header}} | {{Team:Groningen/Notebook/Day/Header}} | ||
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'''Transformation of ''E. coli TOP10'' with BBa_J23109, BBa_J23100 and BBa_J23106''' | '''Transformation of ''E. coli TOP10'' with BBa_J23109, BBa_J23100 and BBa_J23106''' | ||
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* The remainer was centrifuged for 1 min. at full speed and the cells were resuspended in 100ul TY-medium, and plated on TY-Agar-Amp plated. The plates were stored at 37C o.n. | * The remainer was centrifuged for 1 min. at full speed and the cells were resuspended in 100ul TY-medium, and plated on TY-Agar-Amp plated. The plates were stored at 37C o.n. | ||
| - | '''Inoculation of TY-medium | + | '''Inoculation of TY-medium for Glycerol Stocks''' |
* GVP cluster vector BBa_J61035 (2x) in ''E.coli TOP10'' | * GVP cluster vector BBa_J61035 (2x) in ''E.coli TOP10'' | ||
Revision as of 13:16, 6 July 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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Wet
Transformation of E. coli TOP10 with BBa_J23109, BBa_J23100 and BBa_J23106
- BBa_J23109 iGEM plate 2, 2G
- BBa_J23100 iGEM plate 1, 18C
- BBa_J23106 iGEM plate 1, 18O
All dry plasmid DNA in the wells was dissolved in 15ul sterile diH2O, and transfered to 500ul cups.
- Four cups with 50ul competent E.coli TOP10 cells were thawed on ice, and 2ul of dissolved plasmid was added.
- After 30 min. on ice, the cups were put in a heat block at 37C for 5 min. and followed by 5 min. on ice.
- To the cells, 800ul TY medium was added and incubated at 37C for 1 hour.
- 100ul was plated on TY-Agar-Amp plates and incubated o.n. at 37C.
- The remainer was centrifuged for 1 min. at full speed and the cells were resuspended in 100ul TY-medium, and plated on TY-Agar-Amp plated. The plates were stored at 37C o.n.
Inoculation of TY-medium for Glycerol Stocks
- GVP cluster vector BBa_J61035 (2x) in E.coli TOP10
- Terminator vector ? (2x) in E.coli TOP10
- 4.5 ml TY-medium with Amp. was inoculated with a colony grown on TY-Agar-Amp. plates o.n.
- The tubes were put in a waterbath at 37C (150 rpm) and grown o.n.
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