Electroporation of blastema cells

From Tanaka Wiki
Jump to navigation Jump to search
The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.

Procedure

  1. Incubate Blastema Cells 20-30min with 1mM DPBS/EDTA
  2. Prepare 1.5ml tubes with DNA mix (max. 3µg/µl, 0.75-1µg/µl per plasmid). Max. 3µl DNA mixture
  3. Prepare 12well plate with gelatine
  4. Prepare A1 HS Media
  5. Centrifuge cells for 3min at 700g
  6. Resuspend one flask (75cm2) with cells after centrifugation in 160µl Steinberg solution
  7. Add 12µl each of the cell suspension to every DNA mix
  8. Electroporation conditions are: 700V, 40ms, 3 pulses
  9. Add cells to the plate and check after 1 day
Retrieved from "https://wiki.tanakalab.org/index.php?title=Electroporation_of_blastema_cells&oldid=686"