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Lab Aug 31 2009
From 2009.igem.org
Plan
Today: Monday, August 31
- Pick and broth culture 7 samples of K235000 from plates
- Assemblies
K235009=J23102(Amp)+J23032(Amp)+P1010Cm
K235010=J06504(Am)+B0015(AmpKan)+P1010Cm
K235011=K235009(Cm)+K235010(Cm)+P1010Kan
K235012=K235011(Kan)+K235000(Cm)+P1010Amp
- Done already: J23102(Amp) upstream, B0015(AmpKan) downstream, J06504(Amp) upstream, K235000(Cm) downstream
- Digest: J23032(Amp) downstream
- Ligate K235009 and K235010 and transform onto 2 Cm plates each
- Transform CCDB with P1010 Kan from new rehydration onto 6 Kan plates
Tuesday, September 1
- Check plates- if P1010 Kan is working we can continue, if not, we must find a different plan
- Make multiple broth cultures from each P1010Kan, K235009 and K235010
Wednesday, September 2
- Miniprep broth cultures
- Run gels to check for size
- If P1010 Kan is working we can assemble K235011.
- Digest: K235009 upstream, K235010 downstream, and P1010 Kan destination plasmid
- Ligate and transform K235011
Thursday, September 3
- Check plates
- Make multiple broth cultures of K235011
Friday, September 4
- Miniprep broth cultures
- Run gels to check for size
- If everything is working we can assemble K235012
- Digest: K235011 upstream, K235000 downstream(Done), and P1010 Amp destination plasmid
- Ligate and transform K235012
