IGEM team meeting nodes 7/14/14
Jump to navigation
Jump to search
* Call in information:
* \url{https://zoom.us/j/744239720}
* Or, go to \url{https://zoom.us/join} and enter meeting ID: 820 128 495 (Eri's meeting room)
*
* Weekly meeting:
* \url{https://zoom.us/j/820128495} Or, go to \url{https://zoom.us/join} and enter meeting ID: 820 128 495
*
Attendees: (zoom): Rebecca, Advait, Lou, Ahnon, Craig, Ashley, Patrik, Wes (via zoom)
I. Intro/Go around
II. Report Backs on to-dos from last meeting
* **Research Plan**
* Cloning, verification, purification of proteins
* \url{https://pad.riseup.net/p/realvegancheeseexperiments}
* What, where, \& when is our next experiment? Should we sketch out expts week-by-week?
* sequencing and SDS-page on kappa-casein
* Time line: https
* ://docs.google.com/spreadsheets/d/1NOPblWr9\_pIHMst0Nvy\_\_7J\_n97LNwEsz2ujtZ3yFBo/edit#gid=0
* Need to order yeast asap - anyone with connections to a yeast lab, or ATCC account?
*
* Discussion Notes:
*
* how to remove \_\_\_\_\_\_\_\_ from caseins: alkaline phosphatase
* requires alkaline conditions; bufer we use to do that may denature our pH-sensitive protein
* affects tertiary structure
* reversible?
* unphosphorylated kaseins --> micelles
*
* keep phosphorylation and resuspend in Ca2 buffer
*
* purified caseins --> phosphorylated caseins -->
*
* solubalization of a casein: \url{http://www.google.com/patents/US4055555}
*
* measurements of micelle formation
* UV vis
* protein powder
*
* simplest cheese recipe: milk and lemon juice
*
* cheese making should be occuring in parallel with cloning experiments
* waiting on yeast
* ordered strain from Carolina --> should arrive in 1-2 days
* Here's the ATCC strain we want to order: \url{http://www.atcc.org/Products/All/208280.aspx}
* it's URA3-, and also has a protease deficient mutation, allowing beter expression of recombinant proteins
*
* Stanford has largest yeast genetics program -
* contact anyone involved with yeast genetics or preferably the sacromyces genome database
*
* UCSC iGEM meetup: \url{http://2014.igem.org/Meetups/August\_UCSC}
*
* We have full alpha secretion factor from DNA 2.0
*
* Kim paper used same signal sequence for both Saccharomyces and Pichia
*
* next step: sequence the plasmid; we have 120 free sequence reactions; needs to be done
* move on to synthesis
*
* once we confirm the sequence, can design primers and PCR into whatever vector we want
* can add in additional flanking sites conforming to iGEM standards (biobricks)
* 5 genes total; possible to design a single/universal set of primers for all genes?
*
* How do we decide what to order? How do we reach agreements about when and what to spend money on?
* see google doc regarding ethics: \url{https://docs.google.com/document/d/1qKL4x7lOwG9geCdRng1m2FGfE\_\_aZs4gwUGZPIWxvEg/edit}
*
* next experiments: SDS-PAGE and sequencing from clones from human kapa casein cloning (sups and cell lysates)
*
* develop working groups for tackling experiments in parallel at different locations
* want to avoid transporting reagents back and forth; need to double reagent purchase
*
* human cloning started @ CCL, makes sense to keep human in Oakland and tackle bovine experiments @ BioC?
*
* Eric wants nothing except plasmids to leave BioC; same goes for CCL
*
* timeline for experiments; week-to-week schedule?
*
* monthly BioC meetings as an earmark for progress planning
*
* see google doc: Real Vegan Cheese Timeline
*
* **Stretch Goals**
* *Narwahl (first stretch goal) - Check in with UC Santa Cruz - follow up*
* Sending folks to iGEM - what are requirements to go?
* concern: prohibitive costs of iGEM jamboree conflicts with CCL/BioC/open-source values
* suggestion: alternate trip that jives with CCL/BioC ethics?
* another perspective: iGEM jamboree is a valuable (conference-like) experience; how many people on RVC team feel they would benefit from attending but would be unable to raise funds for trip on their own?
* Picchia pastoris
* later in project
* Bioreactor
* later in project
* Other equipment
* particle-size analyzers
* pH meter
* small (20 L ) bioreactor; not industrial scale; useful for projects beyond vegan cheese
*
* **Update for IndieGoGo**
* Narwhal stretch goal + BioCoder - asap
*
* Craig emailed KQED
*
*
* **Contact Food not Bombs for fiscal sponsorship (Craig)**
* Institute for the Future
*
* **Ethical Team: Maria, Craig, Rebecca**
* We are writing up a document using DIYbio guidelines for ethics, creating internal statements for each point that details how we go about fulfilling our ethics.
* \url{https://docs.google.com/document/d/1qKL4x7lOwG9geCdRng1m2FGfE\_\_aZs4gwUGZPIWxvEg/edit}
III. **NEW TOPICS**
* -Cheesemaking
* Process to make cheese, important factors on cheese coagulation, previous cheesemakign expriments?
*
**underlineCONSENSUS DECISION: Do a second fundraiser for those that want to go to iGEM.underline**
***TO DO (071414)***
* 1. Yeast Strain - finish registration on ATCC (Credit refs)
* 2. Next meeting at BioCurious: have a planned experiment ready for the meeting; contact Nicola to coordinate details
* 3. Contact sequentech about sequencing reactions and primer design -
* 4. Modify experiment timeline google doc: add additional sheets to overall document, one for each month, to break monthly calendar schedule into a week-by-week schedule for precise tuning of progress