GenJet Ver.II,LipoD293及PolyJet针对哺乳动物细胞的DNA体外转染试剂,均为您提供两种不同的转染步骤——一般步骤及高级步骤,这两种步骤分别针对不同的哺乳动物细胞。高级步骤主要针对难转的哺乳动物细胞,例如:MDCK,MDA-MB231,Caco-2等细胞。使用一般的转染步骤若是转染效率低于5%,那么使用高级转染步骤则可以将转染效率提升至60%。
如何选择转染步骤则取决于您的细胞特性。对于诸如HEK293,Hela,CHO,3T3,COS,HepG2,LNCaP,PC3,PC12,U2-OS,L929,MCF-7及Huh-7等常用细胞,一般的实验步骤就能得到满意的转染结果。对于诸如MDCK,MDA-MB231,Caco-2,SaoS-2及HUVEC等难转细胞,可以选用高级转染步骤。针对您的细胞,若是您尚不明确选择哪种转染步骤,我们建议您可先尝试一般步骤,如果您使用一般步骤得到的转染效率低于5%,那么您的细胞比较难转,您可尝试高级转染步骤。
Choose a more efficient protocol for your specific cell.
GenJet Ver. II, LipoD293 and PolyJet DNA in vitro transfection reagent all offer two protocols for transfecting mammalian cells--------a general protocol and an advanced protocol. The two protocol are written for transfecting different types of mammalian cells. The advanced protocol which involves a shaving cell technology is for very hard to transfect mammalian cells like MDCK, MDA-MB231, Caco-2, etc. The general protocol usually gives less than 5% efficiency while advanced protocol can get up to 60% efficiency on these problematic cells.
The principle to decide which one protocol is used to 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 which usually gives very good transgene expression while the advanced protocol does not help. 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 followed by incubation of the cell pellet with transfection complex. If you do not know for your specific cells how to choose the more efficient protocol, we suggest you try general protocol first. If you get less than 5% efficiency with the general protocol, then your cell is hard to transfect in nature and you can consider trying advanced one.