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after REs digestion, ligated the insert , but sequencing showed original fragmen


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#1 biomedical



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Posted 28 February 2012 - 03:22 PM


I have a problem which never met before. After digestion of the vector with two different REs ( NotI and KpnI), igated the insert into cloning sites (NotI and KpnI). Transformation worked. However, the sequencing result showed that there are both original fragment (between NotI and KpnI) and insert ( new NotI and KpnI) in the vector.

Can anyone tell me what happened here? I attached the sequencing result with this message.


Edited by biomedical, 28 February 2012 - 03:25 PM.

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