DsRNA Mediated Knockdown of S2 Cells: Difference between revisions

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**Add ~100 uL cells under coverslip to hemocytometer.
**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.
**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 x is number of cells per row
{{math|<var>&alpha</var> x 4 (rows per square) x 10^5}}
x * 4 (rows per square) * 10^5
**Dilute cells to 1^6 cells per mL and seed out in 0.5mL per 12 well.
**Let cells attach for >30 min, then add 0.5mL fresh media with 10 ug/mL dsRNA
**Refeed cells with fresh media and dsRNA daily, typically for 4 days.


[[Category:Cell Culture]]
[[Category:Cell Culture]]
[[Category:Knockdown]]
[[Category:Knockdown]]
[[Category:Drosophila]]
[[Category:Drosophila]]