is denature plasmid undergo transformation???

Cathal Garvey via methods%40net.bio.net (by cathalgarvey from gmail.com)
Wed Nov 30 06:38:10 EST 2011


There are many reasons why a transformation would give fewer colonies
than you expect.

If you have not yet checked the plasmid preparation on a gel, then you
should do so. Spectrophotometer readings are only useful to distinguish
DNA from protein or RNA, they tell you next to nothing about *what* DNA
you have.

-Cathal
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On 29/11/11 16:30, sudheer sangeetham wrote:
> Dear Researchmates
> 
> I isolated plasmid from zymogen plasmid isolation kit. After pellet down
> the bacterial cells, I added 7x lysis buffer(kit reagent) and by mistake I
> resuspend the pellet with pipette( suppose I should not resuspend with
> pipette). But when I measure plasmid concentration it was 500 ng/mico
> liter. I didnt check the plasmid by running agarose gel. One of my labmate
> told chormosomal DNA also could have come when I resuspend with pipette.
> When I did bacterial transformation with this plasmid I got few colonies.
> Could any please tell, did I do any mistake. Shall I do further steps.



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