pCAT Basic cut by Hind III

Damak, Sami damak at kahu.lincoln.ac.nz
Tue Jun 21 05:36:02 EST 1994


In article <CrKC86.3FD at acsu.buffalo.edu> vp23qw83 at ubvms.cc.buffalo.edu (Xin Guo) writes:
>From: vp23qw83 at ubvms.cc.buffalo.edu (Xin Guo)
>Subject: pCAT Basic cut by Hind III
>Date: Fri, 17 Jun 1994 23:39:00 GMT

>Hi, bionetters:

>I cut pCAT Basic (~4.4kb) using Hind III (one Hind III site) and
>got three bands on agarose gel. Compared with lamda DNA/Hind III
>marker, I saw one the 4.4kb cut linear pCAT band and non-cut
>circular pCAT band. But I also saw a band about 8.8kb. The three
>bands seem to have the same amount of DNA. I am new on molecular
>cloning. I have no idea what the 8.8kb band is. Is it possible
>that the band is ligated two linear 4.4kb DNA or something else?
>I isolated the pCAT from E. coli using phenol/choloroform method.

>Xin Guo
>vp23qw83 at ubvms.cc.buffalo.edu

Yes it is possible.  If your sample was left at a low temperature before 
being run, this could have made the sticky ends generated by Hind III 
reanneal.  I have experienced this with other enzymes and have seen bands 
corresponding to 2 and 3 copies in the gel.



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