IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-17: Difference between revisions
DavidRamos (talk | contribs) |
DavidRamos (talk | contribs) |
||
| Line 53: | Line 53: | ||
==Agenda for the next few days== | ==Agenda for the next few days== | ||
We organized a nice little flowchart going over what we want to do today and tomorrow: | We organized a nice little flowchart going over what we want to do today and tomorrow: | ||
[[Image:Agenda_Flowchart_7-17.png | Agenda for 7/17 - 7/18]] | |||
We determined that the Vent or Vent buffer was contaminated with our template, which caused the streaks in our controls. We decided to do another transformation which we will then miniprep and PCR with new Vent and KOD polymerase. | We determined that the Vent or Vent buffer was contaminated with our template, which caused the streaks in our controls. We decided to do another transformation which we will then miniprep and PCR with new Vent and KOD polymerase. | ||
Revision as of 14:12, 17 July 2006
Images from yesterday's PCRs
- Gels run for 30 minutes at 9:30 AM
 Click for legend |
 Click for legend |
 Click for legend |
- Same gels run for an additional 15 minutes at 2:00 PM
 Click for legend |
 Click for legend |
 Click for legend |
Weekly meeting
Once again we received plenty of good feedback from our weekly meeting.
- There was a strong sentiment that we should synthesize just the KaiABC genes, instead of synethesizing complete operons including KaiABC and the promoters we want.
- + We can order the syntheses sooner, without having to decide on promoters
- + There's a strong chance we'll want to experiment with different promoters, so our synthesized constructs should be modular
- - However, we could still design our operons to have digestible promoters\
- + We would have BioBrick parts for the registry
Agenda for the next few days
We organized a nice little flowchart going over what we want to do today and tomorrow:
We determined that the Vent or Vent buffer was contaminated with our template, which caused the streaks in our controls. We decided to do another transformation which we will then miniprep and PCR with new Vent and KOD polymerase.
Incubation
We incubated our 6 colonies from the other plate in order to increase our chances of getting our 3kb Kai insert. After a lengthy diagnosis, we determined that the 3kb segment we have been working with may not be KaiABC, so this will help us cover our bases.
Transformation
We transformed OneShot Top10 compentent cells with our KaiABC construct from the single clonal colony. This is in preparation of another miniprep and PCR with new Vent and KOD.
Western Blotting Protocol
Found some resources on Western Blots, which can be found here.
