Help! low efficiency of transfection
Posted 14 September 2010 - 11:55 PM
I try to transfect PreB cells which are suspension cells using fugene6 and my DNA is EGFP but i got very low efficiency.
I had been varied cell density, fugene:DNA ratio but not thing better.
I usally use Optimem for dilute fugene reagent and incubate 20-35 mins.
My supervisor told me that even summer student can do this transfection at least 50% efficiency.
Should i try to vary an incubation time to up to 2hrs (from a company trouble shooting guide)?
I had email to a couple of company, most of them told me to use electroporation instead but my supervisor had tried with these cell lines before and she told me that more than 50% of our cells will dead.
Posted 15 September 2010 - 12:19 AM
Posted 15 September 2010 - 04:05 AM