what's wrong with my digestion - (Oct/10/2005 )

Hi,

I digested my pututatively recombinant plasmid, which is 3.9 kb pCR2.1 TOPO plasmid with 697 bp insert. Firstly I digested with HindIII at 37 overnight and then digested with BclI at 50 for 2.5 hrs. It 's expected to get 540bp and 4k , or 283 and 4.3 k fragemnt . But actually I got six bands which are 1kb, 2kb,4kb and 4.5kb. What was that and what's the solution?

thanks

forforfor

hi

i think the bands showed incomplete digestion. i suggest you prolong the incubation time, or use less concentration of plasmid, or use more enzyme(volume). or try all. putative transformed plasmid is unpredictable.

goodluck!

thanks. I 'll do that.