IGEM:MIT/2005/Monday 8/15 Meeting Agenda and Minutes

General Updates

• Kate: Are we coming into your lab today to do more cleaning? When are you around?
  • yes, you have a few things left to do (maybe 20 minutes?)....I'm leaving around 3:45 PM.--Kate
• Team: Info on the Endy lab space:
  1. Jen will show those who weren't around Friday afternoon where our materials are
  2. Heather needs to do some lab specific safety training with everyone, ASAP. We'll do it after this meeting, if we can.
• I may be late, though hopefully not absent, for today's meeting. With apologies, Natalie

SubTeam Updates

Format: what is the progress? what are the issues?

Input: Maxine

No updates.

Receiver Head-Unit: Jen M

• Taking over for Jenny, unless someone else volunteers to.
• This week:
  1. Complete construction of scFv test construct
  2. Begin testing
     • expression (cytoplasmic): western blotting. Any favorite protocols? To plan: talk to team mates and lab members. Timescale?
     • fluorescein binding: need help designing an experiment. should have done by end of week.

Promoter.RBS.scFv 
   [[../../JennyN Lab Notes#Jenny/]] See Detailed Notes Here.
    PCR start codon to scFv
    digest Promoter.RBS BB (recipient)
    digest scFvs (suffix donor)
    gel purify 
    ligate promoter.RBS.svFv
    transform
    overnights for miniprep
    identify correct clones (digest), streak plates
    overnights for freezer stock
    freezer stocks promoter.RBS.scFv
   

Receiver/Transmitter 2 - FecA : Annie

• Make FecA- and Fur-

-Redesign and order primers
-PCR 

• FecA Promoter

-Redigest to verify
-PCR to verify
-Construct FecA Promoter-GFP

• FecA

-Look at Kate's site-directed mutagenesis
-Look at past 2 QuikChange
-Redo QuikChange

• PhoA'

-Digest to verify cut site already removed
-2nd QuikChange

• FecR'

-Design internal primers for sequencing
-Make freezer stock and stock of plasmid
-Send to sequence

• scFv

-Add terminal linker by PCRs
-Biobrick
-Verify size
-Send to sequence

Signal Processor: Ray

No updates.

Actuator

No new updates.