plasmid rescue from both sides of PZ

Gerard Manning ger at stanford.edu
Sun Aug 24 21:37:45 EST 1997


In article <v01540b02b025ef4480d9@[140.141.8.58]>, liebl at CC.DENISON.EDU
(Eric C. Liebl) wrote:

>Does anyone know of a restriction enzyme combination that will allow me to
>rescue genomic DNA from the "rosy gene side" of a PZ insert?

We have used both SpeI and NheI in our lab; neither of these cut within PZ
and so rescue both sites of the insert. We usually then subclone the
result into Bluescript, cutting out most of the vector with HindIII (the
vector cuts to two ~7kb bands, leaving the right hand [rosy-proximal] P
repeat with the insert).

If you want to look for more restriction sites, you can download the
vector sequence from FlyBase:
   http://morgan.harvard.edu/htbin-post/moseg-mlink.pl?moseg=ms00000751

   -Gerard.



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