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==Kidney Samples: Nuclei isolation for Drop-seq== *Samples from Andy Ransick (McMahon lab, USC) gently frozen in 1ml cryopreservative: #Nephron Progenitor Cells 250K 15.2wk #Kidney Cortex (dissociated cells) 10M 16.5wk *Checked for live cells using trypan-blue count before isolating nuclei **Cortex - 32% live cells (post-centrifuge) - ~2.5M total **Nephron progenitors - 16% live (pre-centrifuge) - 160K total *Isolated nuclei using NEB and dounce and a single centrifugation step *Sorted on FACS AriaII (SCRM) - nuclei looked very clean with little debris. Sorted G1-S-G2/M populations. **200K for Cortex **38K for the progenitors (used all of the sample - therfore starting with 250K cells is not enough for Drop-seq) *Centrifuged all nuclei (900g 10min) and resuspended in 100ul of 1:10 dilution of PBSE/1%BSA in 1xPBS (for 0.01%BSA) and transferred to Song for Drop-seq *Experiment documented here: [https://docs.google.com/spreadsheets/d/1um-roPyPw80ZX__IE00KONjAMOFMkXWCkgobQ0QdhA4/edit#gid=0 20161114]
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