Electroporation of blastema cells: Difference between revisions
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== Procedure == | |||
# Incubate Blastema Cells 20-30min with 1mM DPBS/EDTA | # Incubate Blastema Cells 20-30min with 1mM DPBS/EDTA | ||
# Prepare 1.5ml tubes with DNA mix (max. 3µg/µl, 0.75-1µg/µl per plasmid). Max. 3µl DNA mixture | # Prepare 1.5ml tubes with DNA mix (max. 3µg/µl, 0.75-1µg/µl per plasmid). Max. 3µl DNA mixture | ||
Latest revision as of 09:58, 25 March 2015
Procedure
- Incubate Blastema Cells 20-30min with 1mM DPBS/EDTA
- Prepare 1.5ml tubes with DNA mix (max. 3µg/µl, 0.75-1µg/µl per plasmid). Max. 3µl DNA mixture
- Prepare 12well plate with gelatine
- Prepare A1 HS Media
- Centrifuge cells for 3min at 700g
- Resuspend one flask (75cm2) with cells after centrifugation in 160µl Steinberg solution
- Add 12µl each of the cell suspension to every DNA mix
- Electroporation conditions are: 700V, 40ms, 3 pulses
- Add cells to the plate and check after 1 day