Blue:RNA-Seq Experiments:02272014

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Revision as of 19:02, 27 February 2014 by >B1lake (Created page with "== Human Nuclei C1 Experiment: 02-27-2014 == == Purpose == *To test conditions for preventing Nuclei clumping during C1 loading == Tests == *Sort 50K nuclei per tube in B...")
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Human Nuclei C1 Experiment: 02-27-2014

Purpose

  • To test conditions for preventing Nuclei clumping during C1 loading


Tests

  • Sort 50K nuclei per tube in BSA buffer
  • Centrifuge 910g at 4C for 10 min
  • Resuspend in BSA sort buffer containing:
  • No supplements
  • 1mM MgCl2
  • 25% Glycerol
  • 10% PEG6000
  • 1mM MgCl2/25% Glycerol
  • 1mM MgCl2/10% DMSO
  • Count using Biorad T20
  • Add 7ul (containing 800 nuclei) to 3ul C1 Suspension RGT
  • Incubate 10 min at RT
  • Centrifuge @910G for 10 min
  • Gently resuspend and add into BioRad slide for counting/sizing