For scRNA-seq: Difference between revisions
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(Continuing directly from the dissociation protocol or from FACS)
(Created page with "= scRNA-seq = ---- <li>(Continue from the [[|dissociation protocol]])</li> *After 30um filter cup, immediately filter through 10um filter cup Resuspend the cell pellet with 15...") |
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= scRNA-seq = | = scRNA-seq = | ||
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<li>( | <li>(Continuing directly from the [[dissociation protocol]] or from [[for FACS|FACS]])</li> | ||
* | *Resuspend the cell pellet with 150-300uL AMEM(0) depending on the size of the pellet. | ||
Resuspend the cell pellet with 150-300uL AMEM(0) depending on the size of the pellet. | |||
*Immediately sort the cells into 1.5mL tube with 700uL AMEM(0). | *Immediately sort the cells into 1.5mL tube with 700uL AMEM(0). | ||
**Normally we use F02 sorter because it has lasers for both GFP and mCherry. | **Normally we use F02 sorter because it has lasers for both GFP and mCherry. |
Revision as of 22:39, 18 July 2022
scRNA-seq
- Resuspend the cell pellet with 150-300uL AMEM(0) depending on the size of the pellet.
- Immediately sort the cells into 1.5mL tube with 700uL AMEM(0).
- Normally we use F02 sorter because it has lasers for both GFP and mCherry.
- Note the FACS count.
- Centrifuge 300xrcf, 20 degree, 5 minutes. Remove supernatant.
- The sample is ready for downstream applications: