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.
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