For scRNA-seq

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.