DsRNA Mediated Knockdown of S2 Cells
From Bridges Lab Protocols
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
<math>\alpha\times 4 (rows per square) \times 10^5 (dilution factor)</math>
