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==Kidney Samples: Nuclei isolation for Drop-seq== *Samples from Andy Ransick (McMahon lab, USC) gently frozen in 1ml cryopreservative: #Isolated Nephrons (intact X8; see attached photo) #Nephron Progenitor Cells #Kidney Cortex (dissociated cells) #Glomeruli (dissociated cells) *Tested nuclei isolation protocol on: #250k Nephron Progenitor Cells - used full protocol with 2x centrifugations (500g, 5min) -> very low number of nuclei that looked good but too low to use #600k Glomeruli (dissociated cells) - Full protocol -> mostly debris that looked like cell clumps and that were DAPI+ #600k Glomeruli (dissociated cells) - Just up to dounce -> same as 2, mostly debris that looked like cell clumps and that were DAPI+ *Did not see any cell pellet for the Glomeruli, maybe something happened during freezing? *For Nephron progenitor cells, maybe the centrifugation was too low (need to use 900g 10min?) *Need to run through my protocol with control cells to make sure it is working...
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