For scRNA-seq: Difference between revisions
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(Continuing directly from Liberase dissociation or from FACS)
| Line 9: | Line 9: | ||
**If there are too many doublets or clumps, better repeat the experiment. | **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 | **If the concentration is too low or too high -> spin down the cells and resuspend in a different volume | ||
*Follow the 10X scRNA-seq | *Follow the 10X scRNA-seq protocol from here, or submit the samples to the NGS facility. | ||
Latest revision as of 22:58, 18 July 2022
scRNA-seq
- 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.