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| * contact PETA and other vegan organization | | * contact PETA and other vegan organization |
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| ---- old meeting ----
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| May 12th 2014
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| = Agenda =
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| == Introductions ==
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| Icebreaker question: How is genetic engineering going to save/destroy the world?
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| == Brief intro to project for new people ==
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| == Announcements ==
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| == Updates from sub-projects ==
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| What happened in the last week? A brief overview.
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| === Fundraising ===
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| === Molecular biology ===
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| * Craig and Juul have construct designs
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| === Cheese making ===
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| * Chr. Hansen wants to know exactly which products we want.
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| === Background research ==
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| == Break out into sub-groups ==
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| Sub-group "break-outs" may have to occur in serial because of overlap of members.
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| Groups should plan next weeks work and assign tasks.
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| We encourage people to meet one on one or in their sub-groups to work during the week.
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| === Fundraising ===
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| === Molecular biology ===
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| Proposed goals for upcoming week:
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| * Finalize construct design
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| * Plan initial experiments necessary to get "band on a gel" verification of casein and kinase expression
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| * Make list of all required reagents, tools and other consumables, including gBlocks and primers
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| * Reach out to companies asking for donated reagents (and plasmids from DNA 2.0)
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| === Cheese making ===
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| === Background research ==
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| ---------- Notes from previous meeting ----------
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| Various intros by lots of people present. Unfortunately no notes for introductions. 22 people present.
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| - Registrations on http://www.meetup.com/BioCurious/events/180642612/ and http://www.meetup.com/Counter-Culture-Labs/events/174105062/
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| - ccl-igem@googlegroups.com
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| - wiki: https://www.counterculturelabs.org/index.php/Vegan_cheese
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| - sci-hub.org
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| * Patrik summarizes the vegan cheese project and iGEM for new people.
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| = Agenda =
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| == Intro ==
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| Patrik
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| What is Casomorphin Casomorphins are peptides, i.e., protein fragments, derived from the digestion of milk protein casein.
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| What are their effects?
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| Do they need to be considered in the formulation of cheese?
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| == How can people sign up for the iGEM list/stay in touch with the team? ==
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| Google Groups Mailing List: ccl-igem@googlegroups.com
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| Sign up at https://groups.google.com/forum/#!forum/ccl-igem
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| == What is the work that needs to be done by this team? ==
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| 1. Fundraising
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| a. Sub-team to take the lead on this? Crowdfunding? Who are we reaching out to?
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| b. Support from corporations? (Mallory can reach out to - All tech in Ky bio ag company, New Harvest, Maria can contact Barracuda Networks) EPA?
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| c. Fundraising goals $3500 fee for registration, materials $5000 Goal low $15k, medium $25k, high $50k
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| d. Ask what crowdfunding we are/will do, crowdtilt has been done, going to restart
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| e. Treasuer to keep track of funds, expenses, and goals with an estimated budget
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| f. Are donation tax deductible? who's 501c3 would do this? 501c3 applied for though CCL
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| g. Teach a class realted to the proejct, all funds going to the project
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| h. speaking at events i.e. First Fridays
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| i. go out to speaking events like first Oakland and talk about project
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| j. Scrappy vegan biohaker mascot - farmer milking a yeast cell with udders for marketing
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| k. organize a GMO conference!
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| 2. Molecular Biology
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| a. synthetic biology services: IDT http://eu.idtdna.com/pages/landing/igem
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| - 25% off an order, up to $800
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| - 40% off gBlocks® Gene Fragments, MiniGenes, or Genes
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| b. What constructs do we want to make?
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| - triple-gene, inducible, construct with promoter-regulated transcription of cheese components.
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| - single constructs that serve as controls +/- expression tags for purification.
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| - kinases needed for the polymerization of Caseino-peptides.
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| c. Kinases and glycosylation - Do we want to co-express anything?
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| d. can we create a construct/work around yes/no flow chart. IE: if construct is created and secretes, then...
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| 3. Cheese Making
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| a. what is the process? Is is the same for synthetic cheese when compared to dairy-derived cheese?
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| b. What are our equipment needs?
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| c. Experiments?
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| - determining the source of fat
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| Experiments:
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| 1. Cheese Track
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| a. Make powdered casein and make cheese from it
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| b. powdered casein with Ca2+ and PO4 to make cheese
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| c. Synthetic Casein + CA2+ to cheese
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| d. synthetic milk to casein
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| 2. Protein Track
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| a. Express various caseings
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| b. evaluate mixtures of synthetic and natural caseins
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| c. optimize casein phophates
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| d. results in syntehtic milk -- make cheese from syntehtic milk
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| 3. Recipe Track
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| a. Evaluate pH History of Water/Pwder Mixture
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| b. Develop recipe for reconstituted milk
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| c. Develop Recipe for Micelle formation
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| d. Develop recipe for cleaving enzyme
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| e. artificial micelles from fat emulsifications.
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| 4. Background research
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| a. Cost of productions
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| b. Are human casains appropriate for adult humans? research about this
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| $83 in donations tonight (Patrik) - where do those funds go? Who holds them?
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| Fundraising;
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| Maria
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| Mallory
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| Patrick
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| Molecular biology:
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| Brian
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| Dimitri
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| Teddy
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| Johan
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| Marc
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| Craig R.
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| Nikola
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| Students
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| Colin
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| Advait
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| Blake
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| Cheese Making:
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| Teddy
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| Maria
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| Brian
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| Della
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| Background:
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| Dimitri
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| Mallory
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| Molecular Biology
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| Genes
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| Full-length bovine κ Casein
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| B genetic variant AAQ87923.1
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| Full-length bovine αs1 Casein
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| Full-length bovine αs2 Casein
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| Full-length bovine β Casein
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| A2 genetic variant for health benefit
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| B genetic variant for better coagulation
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| Full-length human κ Casein P07498.3
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| Full-length human αs1 Casein
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| Full-length human αs2a Casein CSN1S2A
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| Full-length human αs2b Casein CSN1S2B
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| Full-length human β Casein
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| human caseinomacropeptide
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| FAM20C Kinase
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| Promoters
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| 3 promoters (low, mid, high)
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| Selection Markers
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| http://protocols.mmml.nl/protocols/yeast/yeast_selectable_markers.php
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| OriC
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| Purification Tags
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| Constructs:
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| Full length single peptides
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| do promoters as separate biobricks (if needed)
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| have other iGEM projects done protein secretions
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| HIGH Copy Vector
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| Low Promoter
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| SS+
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| Mid Promoter
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| SS+
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| TAG
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| High Promoter
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| SS+
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| MID Copy Vector
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| Low Promoter
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| SS+
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| Mid Promoter
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| SS+
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| TAG
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| High Promoter
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| SS+
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| LOW Copy Vector
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| Low Promoter
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| SS+
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| Mid Promoter
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| SS+
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| TAG
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| High Promoter
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| SS+
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| Database of constructs, sequences and restricitons
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| HiFi Polymerase - PfuUltra
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May 19th 2014
Agenda
Introductions
Patrik: Bio
Jamie: Biotech student at BCC
Rebecca: Graduate student. Synthetic biology, science fiction and history of science.
Shashank: Studying salt tolerance in plants
Advait: Middle schooler interested in biology
Mallory on Zoom
Fran (not on Zoom, only this document): Nursing student learning about biology.
Brief intro to project for new people
wiki: https://wiki.counterculturelabs.org/index.php/Vegan_cheese
Announcements
Updates from sub-projects
What happened in the last week? A brief overview.
Fundraising
Fran: Video script is written and Craig provided some very helpful initial comments. I will give recording another shot in the morning, and then upload a version with no music soundtrack and one with a music soundtrack to solicit further comments. I am committed to getting this recording done tomorrow. Anybody should feel free to take a crack at it, though, I don't want to hog the project.
In any case, once the next iteration of the recording is done, we will still need a lot of input about photos, stock footage, any captions/titles, etc.
Finding good stock footage has been challenging! Easiest route to good footage to layer into the video might just be to take it ourselves with cell phones or other devices.
Molecular biology
Craig presents on cloning.
Many vectors have multiple cloning sites: A series of restriction endonuclease sites.
You amplify your gene using PCR or cut it out of another plasmid, matching restriction sites and ligate it into the plasmid.
Meh, I can't pay attention and take notes. Here's the presentation:
https://docs.google.com/viewer?a=v&pid=forums&srcid=MDQ3ODI0ODE4NDM3NTM4NjIyMzUBMDM5OTQ5MTk5OTAwNDIzOTE2MjcBa0Z0ZTFkd2xycGdKATQBAXYy&authuser=0
https://www.youtube.com/watch?v=WCWjJFU1be8
Cheese making
Miyoko Schinner - vegan chef and TV host, has written books on vegan cheese and has signed up on our meetup list. I'll (Ahnon) contact her directly.... just sent her em via meetup.
= Background research
Break out into sub-groups
IP Issues discussion
Marc: should definitely patent, and do defensive license; willing to write up draft asap
Patrik: fastest way to get most people eating vegan cheese would be patent the heck out of it, show it can be made cheaper than cow's milk, and sell the IP to Kraft. - If that is the *only* issue we care about.
Mallory: if you open everything, then groups with most resources will get the upper hand
Mallory was basically making the argument that big incumbent corps will take what we made and make lots of profits.
Fundraising
Patrik: If we have a fundraiser we really should put a lot of effort behind it.
Ahnon: needs to contact Amyris again
Patrik: needs to contact Bruce German to ask about companies working on breast milk nutritional supplements
Advait: http://www.ncbiotech.org/research-grants/research-funding/biotechnology-research - too late to apply, maybe next year
Advait: http://www.gofundme.com/
http://www.rockethub.com/
http://peerbackers.com/
Jamie and Marc will contact PETA office in Oakland (do they have contact with Patrick Brown as well?P
Shashank: Viz. Amyris, saw Greg Warner at BioCurious today. Shashank - please remind Greg to join the iGEM team! I don't think he' son our mailing list yet. Do you have an email for him? Maybe the one I've been using doesn't work -- Patrik He was away on vacation..i have a netcom.com email.will forward a note to both of you...-s gwerner@ix.netcom.com - yeah, the ix.netcom one is the one I've been using
http://www.gofundme.com/
http://scistarter.com/
https://experiment.com/
Molecular biology
iGEM DNA distribution kit:
http://parts.igem.org/Help:2014_DNA_Distribution
Proposed goals for upcoming week:
- Finalize construct design
- Plan initial experiments necessary to get "band on a gel" verification of casein and kinase expression
- Make list of all required reagents, tools and other consumables, including gBlocks and primers
- Reach out to companies asking for donated reagents (and plasmids from DNA 2.0)
Cheese making
Tasks
juul
- Look more closely at DPL. Follow up with Linda Kahl and Brewster Kahle.
- Follow up with Rusty to see if he can help with video / equipment
- Find the best plasmid for us to order from addgene (but check the biobrick dna distribution list first)
Patrik
- Follow up with Bruce German Re: milk protein companies
Jamie
- contact PETA and other vegan organization