DsRNA Mediated Knockdown of S2 Cells: Difference between revisions
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**Count one row of 4 squares. For cells touching the edges of the square, only count the top and left line to avoid counting the same cells twice. | **Count one row of 4 squares. For cells touching the edges of the square, only count the top and left line to avoid counting the same cells twice. | ||
**Record the number of cells (should be 30-150 cells per row) and Calculate cell concentration, where \alpha is number of cells per row | **Record the number of cells (should be 30-150 cells per row) and Calculate cell concentration, where \alpha is number of cells per row | ||
< | {{math|<var>&alpha</var> x 4 (rows per square) x 10^5}} | ||
[[Category:Cell Culture]] | [[Category:Cell Culture]] | ||
[[Category:Knockdown]] | [[Category:Knockdown]] | ||
[[Category:Drosophila]] | [[Category:Drosophila]] | ||
Revision as of 20:03, 29 September 2009
Materials
- S2 Cells. See Culturing S2 Cells
- dsRNA. See Preparation of dsRNA
- S2 Cell Media. See Culturing of S2 Cells
Protocol
- Take a near-confluent plate of S2 cells aspirate media and scrape into 1 mL of fresh media.
- Add 9 mL of fresh media and pipet to mix.
- Using hemocytometer count cells.
- Add ~100 uL cells under coverslip to hemocytometer.
- Count one row of 4 squares. For cells touching the edges of the square, only count the top and left line to avoid counting the same cells twice.
- Record the number of cells (should be 30-150 cells per row) and Calculate cell concentration, where \alpha is number of cells per row