how to judge the result of BstXI digestion

Peter Ianakiev ianakiev at genome.wi.mit.edu
Wed Feb 13 10:16:12 EST 2002


Hi dear
I don't know this vector, but I guess that the point of using BstX-I is
that both ends after digestion are not compatible. That is also the
reason why they put two BstX-I instead of one. So your test is fairly
simple - if you don't have many colonies after Vector + Ligase control,
then  you have complete digest.
Cheers
Peter

shude yang wrote:

> Hello, Everyone:
>
> I use plasmid pFL61 as a vector to built cDNA
> library.The DNA of pFL61 has two BstXI recognise sites
> between which there are only 20bp nucleotides.
> Everytime I use BstXI to digeste the plasmid pFL61,I
> can not find the difference between the moving
> distance of digested and undigested DNA of pFL61 in
> the agarose gel,and can not find the 20bp-long
> internal BstXI frangment either. Can you tell me how
> to determine whether the DNA of pFL61 is digested.
> Sincerely welcome any instruction .Thank you.
>
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