5 replies to this topic

[Oracle602]

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

[Youlia]

Which organism are you work with?

[Oracle602]

E.coli
Trying to incorporate various environmental resistance genes

[Youlia]

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

[chandima]

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

[peixumol]

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