Hii..I want to transfct H9C2 cells with plasmid having 8.5kb (shRNA plasmid construct).
By which method i can get maximum efficacy.?
You should use Lipofectamine 3000 from Thermo Fisher. I've read reports of large plasmids transfection using it and it also works for H9C2 cells.
https://www.thermofisher.com/br/en/home/brands/product-brand/lipofectamine/lipofectamine-3000.html
Dear Bugga,
I would definitively recommend the Magnetofection technology***. This method was demonstrated to be highly efficient in transfecting any kind of nucleic acid (DNA, siRNA, ODN...) in any type of cells, cell lines as well as primary cells (from neurons (any type) to endothelial cells, chondrocytes, muscle cells...). For a complete list of cells, you can follow the link attached.
This method present a high compaction level of nucleic acid (ideal for large plasmids) and the material used is cell life compatible and biodegradable: toxicity is thus reduced.
I would be very glad to assist you in using this technic; please do not hesitate to contact me via Researchgate or directly at : [email protected].
Good luck in your experiment,
Best regards,
Cedric
*** The Magnetofection technology uses a magnetic field to attract and concentrate complexes of magnetic nanoparticles and nucleic acid onto the cell surface as demonstrated by Grześkowiak BF et al, Pharm Res. 2014 Jul 18. (http://www.ncbi.nlm.nih.gov/pubmed/25033763).
http://www.ozbiosciences.com/module/citationfinder/default
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