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

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Contents

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.

Subteam Updates

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.

ToxR-Will

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

Signal Processing-Ray

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
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