IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-21

Results of Transformation

Literally more colonies than one can count. Prehaps 2uL is the best to use in the future? There are 11 plates, 5 for 36010+36012 and 5 for 4500+36001, and 1 for b0015 in the fridge.

Restreaked 3 colonies from each of 36010+012 and 4500+36001 plates. The 4500+36001 series was called by letters, the 36010+012 plates by numbers, and roman numerals for the b0015.

PCR protocol:

• 94 - 5min
• 94 - 30s
• 55 - 30s
• 72 - either 1min for 4500+36001 or 4min for 36010+012
• Cycle to 2, 30x
• 72 - 10min
• Terminate

Each rxn: 8uL supermix, 1uL VF2 @2nm 1uL VR @2nm

Meeting Notes

• Remove 'a' and 'c' in corner of images
• Remove KaiABC extraction slide
• Simplify synthesis slide (remove advantages, disadvantages)
• Hunt rhino to extinction on slides after first (awesomeness <-> time)
• Simplify Ellowitz plasmid
• Recreate KaiABC diagram w/o extraneous details

Hi Jeff

Slide 2:

Content does not match title – explain; split into two possibly; (“previous work”) - switch order of two? Delete flexible period / cite source – or say possible to mutate kaia – (though push back to later); examples of engineering systems? Fit it to endy’s goal; prehaps how a clock would be usefl to frame it in (prehaps if we had a bioclock…) Erase everytrhing but center traignle

Slide 3:

Simplified View – Reduce text; delete last point?

Slide 4:

Don’t make it orange

Slide 5:

delete this slide but don’t use extraction anyways; use “clone out of organism”

Slide 6:

no need for advantages/disadvantages; prehaps we don’t need a full slide

Slide 7:

Reconstitution of KaiABC instead of “interaction?”; take out the (ip phosphorylated KaiC); cartoon diagrams of kaiABC, same type as intro; prehaps 3 columns?; “KaiC never phosphorylates”

Remove or do something with implication arrows

Number experiments

Slide 8-10: none

Overview of experiment slide: functionalize, emphasize measured quantity

Foreshadow results

Results: construct diagrams, magic

Results

Looks like we got all of what we want :d

grew 30mL of "I" , "1", and "E" in Amp + Kan resistance.

expected size: 857bp

expected size: 857bp, for labeled II different as it is B0015

File:2006-10-21 010+012.jpg

expected size: 3204bp

File:2006-10-21 010+012.jpg

expected size:3204bp, I might have loaded the last lane wrong but still weird band