User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/08/08: Difference between revisions
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# Add the total 600 μL of DNA+Opti-MEM+PLUS reagent+Lipofectamine mixture '''drop-wise''' to each appropriate well of cells. | # Add the total 600 μL of DNA+Opti-MEM+PLUS reagent+Lipofectamine mixture '''drop-wise''' to each appropriate well of cells. | ||
# Incubate cells at 37°C in a CO<sub>2</sub> incubator | # Incubate cells at 37°C in a CO<sub>2</sub> incubator | ||
# | # To reduce toxicity, after 6 hours wash cells once with warm 1xPBS to remove excess DNA-Lipo complexes. Add fresh antibiotic-free growth medium. | ||
Transgene expression should be detectable after 18 hours. | Transgene expression should be detectable after 18 hours. |
Revision as of 12:53, 9 August 2013
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08/08/2013 Gal4-VP64, Cyan Transactivation1 day before transfection:
Transfections > U2OS Flp-in T-REx lines (puro/zeo/blast) + FRT-marked mammalian transfection vector (hygro)
Transgene expression should be detectable after 18 hours.
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