SEQUENCING Large Plasmids

Michelle Larsen ml11 at columbia.edu
Fri Sep 1 10:30:36 EST 1995


I routinely sequence 60-70kb plasmids with no problems.  I use the USB 
Taq Cycle sequencing kit and end label the primers with P-33.  I have 
also sequenced much smaller plasmids (3-25kb).  Let me know if you want 
more details.

Michelle Larsen
ml11 at columbia.edu

On 31 Aug 1995, Steve Wylie wrote:

> Has anyone tried to DNA cloned into plasmids of around 12kb? Presumably 
> one would need to add more template than with a smaller plasmid as the 
> proportion of target DNA to the rest of the plasmid is less. ABI tells me 
> it is better not to attempt it, but I would like to check the site of 
> insertion into the vector. Any suggestions most welcome.
> Steve Wylie
> Murdoch University
> Western Australia
> 
> 



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