Blue:RNA-Seq Experiments:12052016

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Revision as of 20:28, 5 December 2016 by >B1lake (Created page with "===PDX Single-Cell RNA Seq=== '''Meduloblastoma tissue (PDX) from Weschsler-Reya Lab''' *Tissue was flash frozen on 10/19/2016 (3 separate chunks) *Isolated nuclei: :**Remove...")
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PDX Single-Cell RNA Seq

Meduloblastoma tissue (PDX) from Weschsler-Reya Lab

  • Tissue was flash frozen on 10/19/2016 (3 separate chunks)
  • Isolated nuclei:
    • Removed from -80C, transferred quickly to NEB and used scalpel to mash, transferred to dounce
    • 5 strokes with loose pestle
    • 15 strokes with tight pestle -> ice 10 min
    • Filtered 50um filter
    • Centrifuged - re-filtered (30um filter) before FACS -> 5M nuclei
    • Sorted 200K for DROP-seq
    • Froze down remaining unsorted nuclei as potential bulk control


Note: Tissue was very amendable to dissociation