IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

restriction site lost in DH5-alpha

Tracy Aquilla aquilla at salus.med.uvm.edu
Fri Jul 15 12:37:27 EST 1994


In Article <CsopKM.Ars at acsu.buffalo.edu>, vp23qw83 at ubvms.cc.buffalo.edu (Xin
Guo) wrote:
>
>Hi, everyone:
>
>I purified pCAT from JM101. We have made an insert in this pCAT
>between Hind III and Pst I sites. The restriction enzyme digestion
>and sequancing also show right insertion. After transformed DH5-alpha
>cells, pCAT lost Pst I site. Is this a usual result? How do you make
>sure you get the right colonies after transformation (with amp+ 
>resistance)? 
>
>Xin        vp23qw83 at ubvms.cc.buffalo.edu

    How do you know the site was lost, did you sequence this region? How
many bases are missing/extra? This is an unusual result in my experience.
(DH5-alpha is recA minus).
                Tracy



More information about the Methods mailing list

Send comments to us at biosci-help [At] net.bio.net