Blue:RNA-Seq Experiments:12052016
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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