Transformation Efficiency - (Dec/25/2004 )

What factors would you say could affect the transformation efficiency in incorporating supercoiled plasmid DNA into a unicellular eukaryote?
I used the calcium chloride protocol. Temperature of incubation? heat shock? amounts of cells/plasmid used?

Thanks in Advance,
bigO

Which organism are you work with?

E.coli
Trying to incorporate various environmental resistance genes

Hi

There are different methods of E. coli transformation, including heat shock (which could be performed at 37 or 42 C degree) and electroporation.
It also strain and insert dependent, and if you use old cells it might also not work.

Good Luck

hi and happy new yr
will CaCl2 mtd perform better to electrophoration in E.coli transformation. I tried electrophoration and always got blue colonies only

the ligation effiency,according to my experience. the longer time your ligation processes the higher Transformation Efficiency will be! good luck!