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Problem with the transformation


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#1 saad

saad

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Posted 03 March 2009 - 04:36 PM

Hi all
I did transformation with E coli then I did cell cultre.... miniprep...restriction enzyme digest then 1% agarose gel electrophoresis
as you have seen in the gel i repeated this experiment two time one of them duplicate for two insert with same plasmid but I got a strange result as I enclosed in the photo
could you help me find what is the problem? I try to do every thing ideally....
Posted Image
Posted Image
Visit My Website lane 2 and 3 the same plasmid with the same insert lane 4 and 5 the same plasmid with different insert
Visit My Website lane 2 is the same as lane &3 above , lane 3 is the same as lane 4&5 above
"
Sorry I can't upload the photos on this page"
Thank you in advance

Edited by saad, 03 March 2009 - 04:45 PM.


#2 NemomeN

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Posted 03 March 2009 - 07:04 PM

Looks like some nicely supercoiled pBS...it happens from time to time. The other option is re-arranged plasmid which can be seen if you have an unstable backbone....you end up keeping the ori and selection marker and lose the rest.




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