IGEM:MIT/2005/Friday 8/26 Meeting Agenda and Minutes
- Who's leaving? Who's coming back? When
- Jen - leaving today - back when school starts
- Maxine - leaving Tuesday
- Annie - leaving Tuesday
- Will - ?
- Ray - left due to illness
- Jessica - back on Monday
- Semester meeting schedule - Let's try Friday at 5pm. The meetings will necessarily be short. We can always try something else.
- Doing experiment on microscope today
- Is there any access to any other fluorescent microscope? I waited a few days to use it.
- Backtracked a few weeks, decided to not just use the PCR product, but instead put it into vector
- Made new Biobricks (need to be specified) consisting of ATG-scFv (x3)
- To do: assembly with promoter::RBS construct, western blot testing.
- Does someone want to take over? If so we need to talk about it today.
- Construction of FecA Pro-RFP, FecA Pro-GFP
- Failed once
- Did again and got lots of colonies for Feca Pro-GFP, but 2 - controls got some colonies as well.
- -Having Registry assembling as well
- 2nd QuikChange no colonies
- Optimized version of PhoA in Registry, 81% similarity by BLAST
- PhoA Registry has 1 PstI & 1 EcoRI site. PhoA NCBI has 2 EcoRI sites. Diagnosis digestion confirmed this.
- All primers designed for PhoA (PCR & restriction sites removal) were based on PhoA Registry
- Reorder primers to PCR and remove restriction sites to get the "real" PhoA
- 3rd QuikChange resulted in no colonies
- Doing 3-step PCR (running gel to confirm)
- confirming whether or not Linker was added
- ordered primers for sequencing
- ToxR system:
- ctx assembled w/ GFP
- ctx assembling w/ RFP
- FecA system: parallel
- FecA Promoter assembling w/ RFP
- FecA Promoter assembling w/ GFP