IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-28

Western blot images

Western blot images without the annoying rectangle:

JPEG

TIFF

Miniprep of J36335 (KaiA + KaiC), J36832 (KaiB)

We performed 8 minipreps each of J36335 and J36832, eluted in 30 µL EB.

Digest of J36335\S-P (KaiA + KaiC), J36832\X-P (KaiB)

We did 8 digests each of J36335\S-P and J36832\X-P.

Reaction

J36335\S-P

• 7.6 µL DNA
• 1 µL 10x BSA
• 1 µL NEBuffer 2
• 0.2 µL SpeI
• 0.2 µL PstI

J36832\X-P

• 7.6 µL DNA
• 1 µL 10x BSA
• 1 µL NEBuffer 3
• 0.2 µL XbaI
• 0.2 µL PstI

Master mixes

J36335\S-P

• 76 µL DNA
• 10 µL 10x BSA
• 10 µL NEBuffer 2
• 2 µL SpeI
• 2 µL PstI

Aliquot 10µL into each tube.

J36832\X-P

• 76 µL DNA
• 10 µL 10x BSA
• 10 µL NEBuffer 3
• 2 µL XbaI
• 2 µL PstI

Aliquot 10µL into each tube.

Presentation notes

• More pictures in general-- picture of cyanobacteria
• Slide with Western blot: label it with conclusion of experiment
• Cut "Interaction experiments" slide; copy Kai model to results page, delete arrows to show predicted effect
• Conclusion slide, or at least say some closing words before going on to acknowledgements