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Spring Week 2 General Meeting


Spring 2015 General Meeting Week 2

  1. Committee Updates
  1. Administrative
  1. lab cleanout -- 3-4PM Thursday. (April 16)
  1. External Affairs
  1. LA Biohackers Meetup? - Fasih
  2. Would anyone be interested again in a CityLab collab?
  1. Need to fix the powerpoint.  Needs to be more imaginative and needs more “pop” - Fasih
  1. Financial
  1. Spark meeting this past week
  1. Our campaign start time has a problem: it’ll be too late for jamboree registration
  1. We need to either start our campaign earlier, or find other things to pay using our crowdfunded money
  2. mid july start date is fine, need to state that it’s for reagents
  3. ask if there’s an expiration on the account
  1. Infomedia
  1. Tshirt updates
  2. Update Weebly website
  3. Start official Wiki
  4. Help out with Social Media
  1. Slack
  1. set up integrations
  1. Google Docs, Box, etc.
  2. links to weekly meeting agendas
  3. send OWW link to Sri
  1. Danny should make multiple people admins
  1. Lab Management
  1. Vin/Olivia/Julian/Jessica - Let’s organize all of our things in Sri’s space and log everything on the iGEM Quartzy (probably delete or hide the training oligos, we don’t need that anymore).  I had a hard time finding some boxes and stuff, mostly my fault, but having things on Quartzy would help - Fasih
  1. Outreach/Social
  2. Training
  1. Nithin:  We need to get started on developing and streamlining the protocols/lesson plans for next year’s new member training.  Let’s try to hammer it out by the end of the quarter so we can have everything set and ready come recruitment next year - Fasih
  2. maybe start a literature searching class (i.e. fiat lux or upper division) for iGEM members
  1. p/np or graded?
  1. OpenWetWare tutorial (again)
  1. http://openwetware.org/wiki/UCLAiGEM 
  2. add section for sequences that we want to synthesize
  1. Additional information        
  1. QC Bio is supportive of iGEM
  1. $$$
  2. computational capability
  1. Project Updates/Discussion
  1. Silk Genetics
  1. Ordered primers on Thursday, expect to begin PCR and cloning next week.
  1. Silk Functionalization (Fasih, Julian, and Jessica) - Written by Fasih
  1. Albumin Binding Domain Tamura - ordered ABD gBlock with Silver Fusion sites for direct and seamless cloning into the Tamura sequence.  
  2. Julian will be designing and perform the cloning of the ABD in Tamura.  Do we have chloramphenicol or chloramphenicol-plates?
  3. Ordered VF2/VR and SB-prep 2EA/2P-1 primers for ABD/BBc amplification, and linearized plasmid amplification.  
  4. GFP Tamura - Replated glycerol stocks of Tamura.  Grew 500 mL cultures and induced with IPTG overnight.  Both cultures had same conditions, or than being from different platings of the same culture. One culture did not induce, whereas the other one did. Both starter and main cultures grew to OD extremely quickly though, which could be indicative of satellite colony growth on the initial plates.   Need to replate glycerol stocks and observe for satellite colony growth.  
  5. Aliquotted and spun down cell pellets, stored at -20.  
  6. Next week we will perform standard alkaline lysis of the cell pellets, re-suspend in sample buffer, conduct IMAC purification, and hopefully do a coomassie SDS to verify protein presence.  
  7. I attempted a lysis, but used the wrong buffer (KOAc/Acetic instead of 5M KOAc) and degraded the Tamura. Will not make same mistake next time. Such is life.
  8. Need to meet with instrumentation facility and the expression facility soon (next ~1-2 weeks) to get access to microplate reader/miscellaneous equipment, and western blotting modules/imaging, respectively.  
  1. Bee Silk
  1. Began designing biobricks, constructs, and primers necessary for project
  2. looking
  3. into use of 3rd monomer of HB silk, reproducing paper protocol
  4. have designs with promoters, biobricks, expression vectors, gfp, and spycatcher fusion proteins to HB silk
  1. Protein Cages
  1. Meeting with Prof. Yeates on the 22nd
  1. Materials Processing
  1. Dissolving more silk
  2. Dialysis cassettes
  1. 2 mL capacity
  1. ultrapure water
  1. contact: Keegan or Dewal
  1. Open Floor
  2. Advisor Comments

Refer to Sri Kosuri's Page

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