Team:University of Alberta/9 June 2009

Meeting minutes June 8, 2009

The website: Wiki – missing parts of the library (not lost…just can’t get them  ). Fileshare (FTP) – made it in one piece Drupal – that’s gone, but wasn’t a lot on it. The text is saved but not the site formatting. Dean will check it soon. Behaving weirdly, can ssh into it, but it has a different IP address now…JUST WAIT LONGER!

Lab stuff: Starting this week parttimers/volunteers coming in for hours given last week. Several people (who are not at this meeting) still need to sign up for lab times. Clarren had helped with website and logo design but hasn’t responded in a while…will discuss his status next week. Julia will e-mail out updated list of who comes in when. If you said ‘anytime’ give us a specific time, or full-timers may leave early.

Thurs 11-2 Enoch.

Start weekend box for to and from weekend people. Start weekend lab book. REMEMBER TO PUT PLATES BACK in the fridge. –date intials, what it is, concentrations/volume plated. Max drew pretty diagrams Titles and objectives in lab book. Label tops AND bottoms of plates. The lab blackboard gets erased – write down everything important in the lab books, not the board. Put stuff on right side of fridge, second shelf. Be detailed in your lab books

Science teams Reboot/recombineering design: PCRing 48 genes (~2hours to set up) pMM1 – AB and BA cassettes in. Sequencing soon, the maxiprep. Now just mutagenize, then ready to go. Should be done by end of week. Put each gene into just pAB OR pBA (to save reagents for now…can do both later) Primers – basically done. . Looking at the nicking sites – must mutagenize? Could be LOTS of work – 330! Could do Doug’s annealing method instead. Bead: waiting for the beads – give until tomorrow then ‘raise hell’ Will start cloning in selectable markers tomorrow into pAB and pBA Started cloning genes in to pAB. pBA – working so far. Then try PCRing out of them and sticking them together.

Business: redoing business plan (due one week from today). Proposals for funding: sent out today were packages. List of people assigned to different departments was posted on the google groups. Let David know if you can’t do whats assigned. Check the list to see if you’re on it!

Amber will do medical genetics with Julia More people needed for: Comp sci - Eric Bennett and Enoch Engineering - Kalon Graduate studies – Oscar, Justin

Thurs and Fri to tour Jen Hill around the lab – wear LAB COATS and look professional – Max will do it 

Julia will e-mail patch to open Microsoft 07 documents, so you can open sponsorship stuff. Tailor the letter to your study of you can. Definitely spell and grammar check, and make sure the name is correct.

Roundtable: We have iGEM speed code. Spend Deyholos old account first ($400?). Mike apologizes for not being here. Will be here more soon – had to take his daughter to a violin thing, and was in Ontario. Mike will be in the lab.

Strategies for asking for money: Know the science so you can answer!! Because you will be talking to scientists, and they can tell. Explanations:

Reboot and recombineering: want to build minimal genome (yes, really). Build in one block right beside the ori. Then half an m-cherry gene (a red fluorescent protein) will be on either side of our construct. To reboot, you will connect the m cherry halves, pinching out our genome into a new circle. Amp resistance on new construct will select for cells with it. Use the virus Lambda. The original chromosome will cause the cell to glow red. To actually build the construct, use homologous recombination to connect segements that were built on beads. Homologous recombination just needs identical segments on the ends. The strain were using can only recombine the ends, not middles, as the system is weakened.

plasmid: pMM1 = a plasmid, will add the ends to genes so we can build them like lego. Two different versions have the ends on opposite sides, so we can control in which order we link together genes. Use polymerase chain reaction to amplify gene to clone into pMM1.

Beads: paramagnetic bead (imagine magnet inside) with streptavidin attached. Biotin binds streptavidin, and biotin is attached to first gene, with a notI site. You can add pieces on the end of the chain, and ligate them as you go. Add ‘cap’ last with a notI site. Cut with not I and the constructed chain can circularize. Standard high copy origin will be included in fragments constructed on beads. Have SceI site on either side of ori for easy linearization.

PCRing genes: using primers to amplify gene out of genome. Primers have cut sites so can digest and ligate into pMM1

Don’t sound like you’re talking to a 5 year old, don’t convey you think you’re smarter than them.

How it works: Once proposals are ready (due wed, sent to David for approval). Will drop off printed copy to the chair’s desk/secretary. Probably not going to read it, but they can at least read the title. Have delivered by Fri. Call them on Mon to book an appointment. Meet with them. Look professional, business casual. Make sure you understand the science – just ask! Make sure you introduce yourself, be energetic and happy, ask if they’ve read the proposal. Be able to summarize the proposal in a couple minutes. What is iGEM, what we’re doing, how this relates to them. Highlight why our research is important. Ask if they have questions. For science related faculties, ask for contributions of any kind, speed codes we can use, other groups who may want to support us. We’ll take anything! We’re shot gunning…don’t be disappointed if rejected. Ask if they could e-mail the labs in their department, send them our package, ask for any money left on speedcodes. David will give us an estimate later of roughly how much to ask for. Better not to bring up a dollar amount.

Adjournment 