Creating Glowing Green Bacteria for Earth Week

The day before Earth Day, we gathered at Dan's apartment in lovely Park Slope to do our first biotechnology experiment. We used a kit developed for high school students. It had a detailed manual with safety instructions. Basically, E. coli K12 is so safe that you could drink it with no ill effects (although it would taste horrible). We decided to wear gloves anyway, although they were not necessary. The kit is for use in a classroom, with no lab coats, gloves, or eye protection needed. No hazardous chemicals are used, and the E. coli bacteria is easily killed in 10% bleach. So we filled a bucket with it and dumped all our used disposables into it as we worked. We covered the table with plastic, as well as the floor near the workspace. That way we could wipe any spills up, and wipe it down afterwards with the 10% bleach.

To be doubly sure, we obtained the CDC (Center for Disease Control) guidelines for a Biosafety level 1 (BL1) lab and went down the checklist, making sure we complied. Needless to say, we did not start eating the pizza until we were done.

The one time someone (who came late) tried to bring a glass of water into the room, he was yelled at by everyone.

Ellen gave a short talk on biosafety as it applied to this experiment, and a little background on E. coli and the Green Fluorescent protein (GFP) plasmid. The she and Eric, who also has plenty of lab experience, went through the procedure with everyone else watching. And taking notes. And asking questions. And shooting photos. And filming video. It was a bit nerve-wracking, but we got through it.

A little creativity was needed to fulfill the conditions of the experiment. Dan and Ellen tested how long a big glass bowl filled with 42C water would hold temperature when an ice cold object was place in it, and figured out that putting the small tubes in it for 90 seconds would not decrease the temperature by more than one degree. The ice for the chilling of the calcium chloride transformation tubes was put through a blender to chop it finely enough. Other than that, the kit was pretty complete. We probably will do this experiment again at some point with greater participation, after everyone has had a chance to do some "dry runs" with tubes and vials containing water, and plates with nothing on them, to get used to clean technique.

At the end, we had 3 plates because we ended up dividing one of the plain Luria Broth (LB)-containing plates in half. One the LB plate we put a line down the middle, and put the E. coli + plasmid on one side and the E . coli without plasmid on the other. Each of the two LB + ampicillin (the antibiotic which will kill bacteria that did not take up the plasmid) plates got either the E. coli + plasmid or the E. coli without plasmid. If we are successful, then the ampicillin plate with the E. coli + plasmid should sprout little green friends in a couple of days...

Here's video (Part 1) of the evening:

(if you can't play this, go to the YouTube version)

April 15th Meeting - Well, here we go...

We've been contacted by four reporters in the last three weeks. We knew it was bound to happen sooner or later- DIYbio is a hot news topic at the moment because not much has been written yet and there's such delicious potential for sensationalism inherent in the topic (Amateur biologists wipe out Cleveland!) Too bad this came so soon after we formed the group- we were hoping that we'd be incorporated as a nonprofit and have a written set of safety policies set up before we were forced to deal with this. But if you refuse to talk to them, it looks like you are trying to hide something, which we emphatically are NOT, so we welcomed a New York Times reporter into our midst last night, amid the high-volume chatter, clinking dishes, and pirogies of Veselka Restaurant in the East Village.

Song, Daniel, Russ and Ellen were there. Three new folks showed up: Eric, who has been communicating with us on the Google site for quite awhile, and is a molecular biologist. Nurit, who does bioart and is currently working with a group at MIT. Meredith, whose background is computer science and is fascinated by the concept of biological machines. It was great to see the ideas flying as the combination of everyone's divergent skills resulted in some really cool things we are going to follow up. We'll describe some if them in more detail in later posts as soon as we flesh out the details.

Our agenda was to finalize a mission statement for the group, to address what we think the group needs to do in order to be transparent, and to address safety issues and requirements for operating a space where labwork is performed. We were somewhat thrown off schedule by the time needed to communicate to the reporter our goals and hopes for this group, but we still were able to achieve quite a bit.

A draft mission statement was passed around, and we will post it as soon as it's finalized. From what we've read, this statement can be fine-tuned if necessary after the group incorporates, so we just want to make sure we've got the right concepts in there for the initial version. This is the first step to becoming a nonprofit, a step which we feel will make it easier to achieve our goal of productive, socially meaningful scientific research. Russ volunteered to produce the first draft of our articles of incorporation. They'll be posted in the File section of the Google group site.

In terms of transparency, we agreed that it would be useful to keep more formal meeting notes in the future. For now, we are posting in this blog and also keeping track of who attended each meeting. And we'll continue to let reporters observe our meetings. We fervently hope that they don't edit out the hard work, deep thought, and effort that's gone into ensuring this group operates safely at all times.

As we contemplated how to create a safe environment for amateur labwork, several good ideas surfaced. Anyone wanting to do wetwork in our space will have to provide evidence of training (such as a degree in bioscience) or undergo training by members who possess these skills. Ellen agreed to draft a safety training procedure. Dan suggested we limit our initial wetwork to experiments that have already been vetted for safety, such as those from the curriculum of the Dolan DNA Learning Center at Cold Spring Harbor. The "green bacteria" transformation experiment we are going to try at our next meeting is actually one of those.

Since high school students have traditionally been part of iGEM teams, we discussed what our requirements would be for participation of someone under 18. Sponsorship by an existing adult member is a possibility, with the understanding that the adult member would commit to being present during wetwork. We also need to have some sort of liability waiver for all members, like what you sign when you join a gym or rock climbing club.

Space, and the procuration of it, was also a hot meeting topic. The bottom line is, we need to have a monetary commitment from the members in order to support the space. So far, we need a few more who can pony up at least $50 per month in order to support this goal. Shared or part-time space will soon become impractical if we start doing wetwork, so we are talking about a lease, and in an industrially-zoned building. Nurit and others will be exploring options.

As usual, we overstayed our welcome at the restaurant table- the waitstaff was changing shifts and our waiter wanted his tip- so we parted after about 2 hours of fruitful discussions. Next week we meet at Dan's for some more hands-on science. Stay tuned for the video!