Ns125:LabNotes/2015-12-19

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  • DropSeq on Epi Library --> input cells @ 80 cells/uL since not many input cells
  • Do RNA fragmentation + standard rxn + bulk ctrls
  • For RNA fragmentation, use 5 units RNase III in 200 uL total RNA @ 37 C for 4 min with rotation
  • Prepare DICER1 & T2 bulk ctrls with same protocol
    • 250 ng RNA for each bulk sample + 50 uL RT mix + 1 uL polyT primer, incubate samples in thermocycles @ 25 C for 30 min, @ 42 C for 90 min
    • Bead purifcation @ 1.8:1 ratio