Cluzel Lab Notebook
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Glycerol Stocks for MG1655 strains
Forgot to take out the cultures on Friday, so doing this again today.
Reminder: Take culture tubes out of shaker at ~6:30 and put stocks in glycerol! (done)
I get the stage at ~1:30. I will pour 2 featureless molds and try transferring again, gently, to see if we get a clear spot boundary upon transfer.
Result: 5% agarose is extremely bright in GFP field, but no cells were seen in phase contrast. Perhaps more 'wetness' is required to transfer cells.
Note: see below: 5% agarose seems to work fine, no idea why GFP field was so bright before; probably bad settings on the scope
Spot cells directly on gel surface with crimped 30 gauge needle dipped in high-OD cell solution. Trying with both 5% and 3% agarose. Spot cells on slide (w/ needle). Trying with both 3% and 5% agarose. Using GFP cells grown up earlier today.
It is generally hard to find cells if they are only in one location. Features are difficult to locate and so it is difficult to keep the agarose surface in focus. It took me about 10 minutes to find the cell patch on 5% direct application the first time, and another 45 minutes to locate it again!
Images are saved as 3-3* in the ~/dbg dir on the camera computer.