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**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 x is number of cells per row{{math|<var>&alpha</var> x * 4 (rows per square) x * 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:Knockdown]]
[[Category:Drosophila]]