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 x 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 | ||
<pre>x * 4 (rows per square) * 10^5</pre> | <pre>x * 4 (rows per square) * 10^5 (dilution factor) = concentration of cells/mL</pre> | ||
*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]] | ||