pACT2 cloning difficulties???

Riekeltje Koedood rkoedood at bio.bu.edu
Mon Aug 12 15:55:33 EST 1996


Hi All

Recently I performed multiple cloning steps using pACT2 from the yeast 
2-hybrid system, and got very confusing results. Restriction analysis of
the resulting clones does not make sense when comparing it with the maps
I have (got the complete sequence of pACT2 out of GenBank).

Have any of you had such troubles? Specifically: cutting with HindIII-Nco I
or Sal I-Nco I give bands that don't fit the maps at all.

Anyone found the maps to be wrong (though I don't think this is the problem,
rather that something wenawry during cloning).

Any advice/suggestions/comments are welcome.

Thank you.

Marieke



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