IGEM team meeting notes 7/14/14

Agenda 07/14/2014

Call in information: https://zoom.us/j/744239720 Or, go to https://zoom.us/join and enter meeting ID: 820 128 495 (Eri's meeting room)

Weekly meeting: https://zoom.us/j/820128495 Or, go to 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 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: 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: 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: 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: 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. 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?

CONSENSUS DECISION: Do a second fundraiser for those that want to go to iGEM.

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