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Transformation Efficiency


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5 replies to this topic

#1 Oracle602

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Posted 25 December 2004 - 06:59 PM

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

#2 Youlia

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Posted 26 December 2004 - 05:39 AM

Which organism are you work with?

#3 Oracle602

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Posted 26 December 2004 - 10:17 AM

E.coli
Trying to incorporate various environmental resistance genes

#4 Youlia

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Posted 27 December 2004 - 05:27 AM

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

#5 chandima

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Posted 31 December 2004 - 09:09 PM

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

#6 peixumol

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Posted 02 January 2005 - 11:22 PM

the ligation effiency,according to my experience. the longer time your ligation processes the higher Transformation Efficiency will be! good luck!
paul in NanJing China
God helps them who help themselves!




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