Blue:RNA-Seq Experiments:04252017

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Kidney Samples: Nuclei isolation for Drop-seq[edit]

  • Samples from Andy Ransick (McMahon lab, USC) gently frozen in 1ml cryopreservative:
    • Kidney Cortex (dissociated cells) 5M 14.4wk
  • 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.
    • 2x200K nuclei post-sort
  • Centrifuged all nuclei (900g 10min) and resuspended in 20ul of 1PBSE/1%BSA and transferred to Song for Drop-seq
  • Experiment documented here: [1]