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...
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)
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