IGEM team meeting notes 7/28/14

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TODO 7/28/14: Ben (bottomliner) in Quartzy (https://www.quartzy.com/); Ahnon; Matt? Johan; Craig; Ben Patrik has jewelery; need to update IG Ben has camera will photograph afterwards One person from Brazil has selected the t-shirt perk, but has not added $10 for shipping Thiago Penaforte St 697 São paulo, 02318260 Brazil Need more people to update Indiegogo - Advait Need someone to take over the financials - David (bottomliner); Ahnon; Antony Evans might have recommendations Need fiscal sponsors - GenSpace, IFF (Eri), FNB (Gabby)- Ahnon will talk to them Craig - check in re: donations Wiki login problems - Marc (bottomliner); Stan Patrik will send out graphics (done) Ashley can bring 2 test tube racks to CCL Wednesday if still needed Reuters (7/29): Craig; Jamie; Marc; Ben Collect all papers relevant to experiment, and give access to everyone (juul)

  • Solicit questions for ask a biohacker and tell us why we're wrong (juul)
  • Research protein purification methods (juul)

Who can take charge of planning SDS-PAGE gels? Josiah at BioC - Ashley will em him. Rachel has some experience. Happy to help. Who can take charge of planning yeast transformation? Wes, David; Who can take charge of designing DNA for remaining genes? Are we agreed on adding BioBrick prefix/suffix outside the SapI sites for Electra? redesign Ahsley, Marc will look at Cathal's; Patrik Rachel - primers; sequencing

Agenda: 07/28/14 Call-in info: https://zoom.us/j/820128495 Or, go to https://zoom.us/join and enter meeting ID: 820 128 495

I. Intro/Go around (5 min - start on time!) Meeting Attendees: Zoom: Advait, Rachel, At Omni: Matt, Brett, Lou, Ben, Ahnon, Jamie, Patrik, Stan, Johan, David, Wes, Ashley, Shannen, Danny

Note takers this week: Ahnon Facilitator: Lou

II. Report back to-dos from last meeting (10 min) TO-DO list from last week:

  • Collect all papers relevant to experiment, and give access to everyone (juul)
  • Solicit questions for ask a biohacker and tell us why we're wrong (juul)
  • Research protein purification methods (juul)
  • Organize / set up inventory - Ben (bottomliner) in Quartzy (https://www.quartzy.com/); Ahnon; Matt? Johan; Craig;

III. IndieGogo progress (10 min) Need more updates! Update on 3D printed jewelry Patrik has jewelery; need to update IG Ben has camera will photograph afterwards Update on next stretch goal soon-ish - bioreactor More Ask a Biohacker videos One person from Brazil has selected the t-shirt perk, but has not added $10 for shipping Thiago Penaforte St 697 São paulo, 02318260 Brazil Need more people to update Indiegogo - Advait Indiegogo will waive fees if 501c3 from the beginning; not clear if possible after initiation Tax implications Need someone to take over the financials Need fiscal sponsors - GenSpace, IFF (Eri), FNB (Gabby)- Ahnon will talk to them who can take charge of sorting out financials? Contact Food not Bombs for fiscal sponsorship (otherwise Institute for the Future) Figure out tax consequences (compare notes w Antony Evans? ask IndieGogo?) Set up separate PayPal and/or bank account for Real Vegan Cheese? Paypal $ needs to be moved over to bank - Ahnon & Ashley General financial - David (bottomlining), Ahnon, Antony Evan might have recommendations Indiegogo will waive fees if 501c3 from the beginning Patrik still can't log in to wiki (mw sends new pw; allows setting new pw then gives error ("There seems to be a problem with your login session; this action has been canceled as a precaution against session hijacking. Go back to the previous page, reload that page and then try again.") - anyone else? (Revisit when Marc is available); , Stan will help Does everyone have access to wiki, file share?

V Experiment planning (20 min) Who can take charge of planning SDS-PAGE gels? Josiah at BioC - Ashley will em him. Rachel has some experience. Happy to help. Who can take charge of planning yeast transformation? Wes, David; Who can take charge of designing DNA for remaining genes? Are we agreed on adding BioBrick prefix/suffix outside the SapI sites for Electra? redesign Ahsley, Marc will look at Cathal's; Patrik Should we order one more single gene (bKcasein for BioC?), or plan our big order? Where do we order from, and what do we have? Need to collect info on accounts at Sequetech (free sequencing!), IDT (DNA synthesis, incl. big iGEM discount), Teknova (donating plates & media), others (get Sigma account for CCL?) Teknova is sending plates and media but we need to pay shipping Need to get info on donations from Craig Should we appoint a few people in charge of ordering? Johan, Craig, Ben (quartzy hass ordering tools!), Ashley Have human kappa casein Rachel: have insert for the vector; She is order the primers; found the reverse primer that anniels; costs $15 for ....; sequencing will be back w/in 24hours;


VI Casein Powder experiment (remainder of time)

References: Solubilization of casein http://www.google.com/patents/US4055555 Chandrapala et al. Dissolution and reconstitution of casein micelle containing dairy powders by high shear using ultrasonic and physical methods. http://www.sciencedirect.com/science/article/pii/S135041771400131X

Parameters to vary: pH, Ca, T, sonication/shear test using chymosin, and settling over time

I propose we do all this in a bunch of test tubes, so we can vary as many parameters as possible. Do we have a couple large test tube racks? Ashley can bring 2 test tube racks to CCL Wednesday if still needed

Concentration of casein powder: 15g/100ml? (Chandrapala: 10g/100ml, patent: 180 to 240 g/l) Bodytech 100% Casein Powder - unflavored Approximately 80% protein by mass (24g/30g) Approximately 7% sugars by mass (2g/30g) Protease content, 1mg/g We will need to remove the protease before hand, suggestion is to acid wash.

Let casein powder hydrate ("age") for 10-30min before adding additional solubilizing agents (patent). Possibly after or during sonication or shear blending to disperse powder

sonication vs shear blending vs stirring 2 level of temperature: RT vs. 60°C (mentioned by Chandrapala; patent says "preferably in the range from 20°C to 70°C") no additions vs increase pH to 7.0 vs add citric acid to pH 4.6 then KOH to pH 6.8

-> 18 test tubes total

How do we test these? Chymosin treatment and letting the casein powder settle out have both been suggested. Should we split each test tube in two, and try both? If so, we'll need 36 test tubes total!

Reuters: Craig; Jamie; Marc

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