For scRNA-seq

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scRNA-seq


  • (Continuing directly from Liberase dissociation or from FACS)
    • Resuspend the cell pellet with 50-100uL AMEM(0) depending on the size of the pellet.
    • Count the cells using a hemocytometer.
      • Standard protocol: 5uL cell solution + 5uL trypan blue, count the cells in 5 blocks
    cell number/5 x 20 = cell number/uL
      • Ideal concentration is between 800-1200 cells/uL
      • If there are too many doublets or clumps, better repeat the experiment.
      • If the concentration is too low or too high -> spin down the cells and resuspend in a different volume
    • Follow the 10X scRNA-seq protocol from here, or submit the samples to the NGS facility.