- make a solution of 1.25 g LB broth in 50 mL water
- Cap with foil
- Autoclave solutions for one hour
- Let them cool down then at the end of (i didnt think about this earlier but they may have to cool longer than that so we might not even be able to do the next step which means no bacteria this week. ask dr. miller if she thinks its doable or if the broth will be too warm)
- fill a bucket with ice and take it upstairs with you
- go upstairs to costanzi's lab (go in the last door on the left on the third floor if coming from our class lab) and go to the next/second row (to the right) and go to the very back against the window and open the freezer on the left (its a long horizontal freezer). Then on the left hand side of the freezer look for a little box labeled hartings competent cells. Look for the ones labeled BL21 or something like that. You have to limit how long you keep the freezer open because it will hurt the cells and the freezer will start beeping loudly. Put them in the ice bucket. Light a flame (go over to the first row when you walk in everything should be set up over there). Use a pipet tip to scrape a little of the bacteria off and drop the whole thing into the flask. recap and put it on a shaker at 37 degrees. (the speed to shake at should already be set)
last time we did fluorescence but we had to make a concentration curve to figure out the concentrations of our dye. while the things are autoclaving could you graph the data (its in dropbox, fluorescence, april, kln) and try to figure out the area under the curve. theres 4 data points, i numbered them so theyre easy to find.
- Add data and results here...
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