Wednesday, November 10, 2010

protocol for ROX1-YFP induction

put 1 matchhead cells from freshly streaked cells into 100ul
put 50ul of this into 10ml deoxygenated media and allow to grow at 30' for ~20h
prepare microscopy plate (treat with conA). place plate on microscope and remove media from target wells. (note: it takes about 5m for the plate to stop moving once you put it on the plate-holder--you should have it stop moving before adding cells, so you can take high quality images immediately after plating.)
take 2ml and spin down in microcentrifuge tube. resuspend in SDComplete.
plate and wait ~1-2m until cells adhere to slide at appropriate density.
begin time-course

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