Blue:RNA-Seq Experiments:12022013
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Human Nuclei C1 Experiment: 12-02-2013
- FACS Sort
- DAPI stained human nuclei from Yun:
- Added ~1ul DAPI just prior to FACS sort on the Aria II (SCRM)
- Nuclei were maintained on ice
- Added ~50ul of sort buffer containing 0.1% BSA to low-bind 1.5ml eppendorf tube for collection
- Gated nuclei on DAPI positive and shifted gate slightly along the FSC axis to exclude the smallest nuclei
- Sorted 150,000 nuclei into the collection tube (final volume of ~1200-1500ul)
- Maintained on ice. Centrifuged 2000rpm at 4C for 10 minutes. Removed supernatant down to ~50ul. Resuspended pellet (not visible) using p1000 by pipetting about 5 times.
- Counted nuclei using BioRad automatic cell counter: 1480 nuclei/ul
- DAPI stained human nuclei from Yun: