DIYbio 3 - Gel Electrophoresis
Thanks to everyone for a great meeting: Michael, Jason, Sophia, Benny, Topher, Ricardo, Alex, Alec, nublabs, and everyone else.
Overview:
DIYbio 2 & 3 were focused on gel electrophoresis - we started by researching all the amateur gel protocols we could find online (notably the MacGuyver Project and the MAKE protocol, the latter of which we decided to focus on) and making a shopping list of materials we would need to build the complete gel apparatus, make a gel, and stain DNA. We got Agar agar powder from a health food store on ebay and Potato Dextrose Agar and sybr green from a lab equipment reseller on ebay.
Basically we over-stained the gels with methylene blue and couldn’t visualize anything because the entire gel was a solid blue.
Also, we used metals that were way too reactive as electrodes in the gel box and the redox reaction that they enjoyed probably ruined the voltage field across the gel.
That said, I think we accomplished a lot, learned a lot, and somehow managed to do everything the protocol required in a spontaneous, parallel way - to me, that was the best part about the event. I remember stepping back at one point and being amazed at how well everyone was working together without any central plan: Topher and Jason were improvising with the stovetop and microwave to boil the agar, Sophia and Mike were cutting up charlie cards to make gel combs, and Benny, Ricardo, and the nublabs crew were constructing and taping gel trays and boxes. It was awesome.
What I learned:
1. We were not as rigorous with the materials as we needed to be. One of our main problems was getting the concentration of the gel just right, which was difficult because the packages for our agar didn’t list the original concentrations. In general, we followed the MAKE protocol too specifically while using supplies that came with ambiguous descriptions at best. DIYbio protocols we develop should be designed to help the user understand the basic principles of the operation in a way that enables them to improvise successfully with non-standard materials.
2. We should have had a solid understanding of how to dispose of all the materials involved before beginning as well as exactly what the potential risks involved were (very low - probably the biggest risk was spilling molten agar) and what to do in case something went wrong.
3. It would have been better to have built all the equipment in one meeting and then used it in a second. Trying to do both was a little too long.
Next week we are going on a tour of the Boston FabLab, courtesy of our friends at NubLabs - meet at the FabLab at 7:00pm on Thursday, 17 July 2008.
See you then!
Mac
p.s. go comment on the flickr photos!
p.s.s cross-posted to diybio blog: http://blog.diybio.org/2008/07/diybio-3-gel-electrophoresis.html
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