Choosing the most efficient protocol for your specific cell typeGenJet Ver. II, LipoD293, and PolyJet DNA in vitro transfection reagents all offer two protocols for transfecting mammalian cells- a general protocol and an advanced protocol. The two protocols are written for transfecting different types of mammalian cells. The advanced protocol, which involves utilizing cell shaving technology, is for difficult to transfect mammalian cells such as MDCK, MDA-MB231, Caco-2, etc. The general protocol usually gives less than 5% efficiency for these cell types, while the advanced protocol can yield up to 60% efficiency for these problematic cells.
The principle to decide which protocol to use for your cell is highly
dependent upon the nature of your cell. For commonly used cells like
HEK293, Hela, CHO, 3T3, COS, HepG2, LNCaP, PC3, PC12, U2-OS, L929, MCF-7 and Huh-7, be
sure to use general protocol, as it usually gives good transgene expression. For very hard to transfect
cells like MDCK, MDA-MB231, Caco-2, SaoS-2 and HUVEC, follow the advanced
protocol by trypsinizing these adherent cells first, then by incubating
the cell pellet with the transfection complex. If you are unsure of which protocol to use for your cells,
it is recommended to use the general protocol as a starting point. If you get less than 5% efficiency when using the
general protocol, you are encouraged to try using the advanced protocol instead. |