IGEM:MIT/2005/Friday 8/26 Meeting Agenda and Minutes

General Updates

 * 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.

Input-Maxine

 * Doing experiment on microscope today
 * Is there any access to any other fluorescent microscope? I waited a few days to use it.

Head Unit-Jen

 * 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.

FecA-Annie

 * 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


 * PhoA
 * 2nd QuikChange no colonies
 * Problems:
 * 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
 * Solution:
 * Reorder primers to PCR and remove restriction sites to get the "real" PhoA


 * FecA
 * 3rd QuikChange resulted in no colonies
 * Doing 3-step PCR (running gel to confirm)


 * scFv-Linker
 * confirming whether or not Linker was added
 * ordered primers for sequencing

Actuator-Jessica

 * ToxR system:
 * ctx assembled w/ GFP
 * ctx assembling w/ RFP


 * FecA system: parallel
 * FecA Promoter assembling w/ RFP
 * FecA Promoter assembling w/ GFP