I want to double transfect two plasmid constructs in SHSY5Y cell line. It is very difficult to perform as I have less than 10% transfection efficiency.
Which is the best method to achieve?
Hi Girish,
First you need a good transfection reagent, you can maybe try our Viromers, see attached file, from page 3 you have plasmid data coming from users.
Second, you need to be sure to use the right protocol. With 1 or several plasmids, it's important to think about he final amount of DNA you transfect and to use enough reagent. What did you try, with which reagent?
Please, give us some details to see if it's possible to help you further.
Best,
Sandra
For cell lines that are difficult to transfect, consider a lentiviral transfection. It's stable and can give much better efficiency.
I can confirm best results with Lonza Nucleofector technology. You can get without any further optimization around 80% transfection efficiency. Please see the attached protocol.
Hi Sandra Lagauzère,
So far I have tried Jet prime, Fugene and Electroporation using OptiMEM. In all the cases, Single plasmid transfection was efficient( more than 60% ) but double transfection is really difficult (less than 10%). And thanks for your suggestion, Hope it works out well.
Hi Peter, Thanks for your suggestion. I will give a try as we have Nucleofactor Kit SF already. I hope it works for double transfection( one DNA in pMAX GFP and other in pMAX mcherry)
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line