Both transfected cells and Cellfectin II Reagent alone negative control cells are dead within 36 hours. What could cause this?
There are several possibilities:
- Using media containing antibiotics during transfections.
- Plating cells at too low a density: We recommend at least 70% confluence.
- Using cells at too early a passage: We recommend growing cells for at least 5 passages before using them for transfection.
- Contamination because of no pen/strep after the transfection: After 5-8 hr incubation with the transfection mixture, remove the mixture and add antibiotics containing media/well.
Find additional tips, troubleshooting help, and resources within our Transfection Support Center.
My transfected cells are floating up and look dead, but the Cellfectin II Reagent alone control plate is fine. What could be happening?
This may be due to contamination or cytotoxicity from the bacmid prep. Make sure to include a negative control that is the bacmid only without Cellfectin II Reagent. Additionally, use the PureLink HiPure plasmid prep kit, not the silica-based miniprep kit for bacmid prep.
Find additional tips, troubleshooting help, and resources within our Transfection Support Center.
I accidentally left my lipid reagent at room temperature. Can I still use it?
Yes, all of our lipid transfection reagents are stable at room temperature for months.
Find additional tips, troubleshooting help, and resources within our Lipid-Based Transfection Support Center.
What is the difference between reverse transfection and forward transfection? What should I use?
In forward transfection, cells are seeded to appropriate confluence or cell density in wells or dishes, and the lipid-DNA complexes are added the next day. In reverse transfection, the transfection complexes are prepared inside the wells, after which cells and medium are added. Reverse transfection is faster to perform than forward transfection, and is the method of choice for high-throughput transfection. For non-high-throughput transfections, generally forward transfections have better efficiency for most cell types.
Find additional tips, troubleshooting help, and resources within our Transfection Support Center.
Is there a place where I can find references from other researchers who have used your transfection reagents?
Visit the product page for each reagent type and you will see a list of references at the bottom of the page. A table that lists specific cell line references is also accessible. We also recommend www.highwire.org as a search engine to find a large selection of up-to-date research articles using our transfection products. Simply include the name of the transfection reagent and your cell line/application of interest in your search criteria.
Find additional tips, troubleshooting help, and resources within our Transfection Support Center.
