User:Daniel Goodman/Notebook/Cluzel/2010/02/26

Cluzel Lab Notebook Daniel Goodman

Main project page Previous entry      Next entry<sitesearch>title=Search this Project</sitesearch>

Glycerol Stocks for MG1655 strains 2 culture tubes 4 ml LB in each 4 ul of Amp in one (for GFP strain) took one average-sized colony from each put in 37°C shaker (at 12:15 pm) Note: I hope I didn't acidentally take from the other tube that was sitting next to mine by the burner; it was Jeff's, looked like LB but it might have had an AB in it. Not too worried though, as long as the cells grow. Retry molds again Will pour 2 molds using silicon chips to generate features, so it will be easier to focus. This must done in the humidity and temp. controlled room so features generate nicely. I am worried about desiccation of the agarose, so I'm going to keep it in the Humidity and Temp controlled room (HTC room) for as long as possible, and get the imaging equipment ready to see if the gel still distorts like on Wednesday. Jeff thinks that using the cover slip on top might not be necessary, but as long as it is placed on top carefully it should not 'squish' the features and it will help reduce moisture loss once the device is out of a high-humidity environment by limiting exposed surface area. There will be no exposed surface area in the actual device, so this hopefully will not be a problem. Make agarose as normal: 5 mL LB, 0.150 mg agarose powder. Vortex in tube, heat at 80°C for 15 minutes. (put in at 12:30) Clean chips in DI water bath (80°C) in flask for 5-10 minutes. (put in at 12:32) After agarose is hot, pour into mold in the HTC room, wait 1 hour for features to form. Put cells on glass slide right before agarose is done setting in normal conditions, move into HTC room and remove from mold and put on top of cells immediately. Cover with square cover slip to maintain moisture.