Lab Aug 31 2009

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