Hi,

I double-digested a plasmid to obtain a 2.5kb DNA fragment that I want to insert into a larger plasmid (11.7 kb). However, I failed to see that my double digest also yields a 250bp fragment with the same restriction sites. Now, both are in the ligation reaction, and I am wondering whether the 250bp fragment will completely outcompete the 2.5kb fragment, or basically what the chances are of getting the 2.5kb fragment inserted despite the 250bp fragment's presence in the ligation reaction.

I ran a diagnostic gel of cut vs. uncut plasmid, and the 250bp band is visible, however much weaker than the 2.5kb band; but this is probably to be expected from the size differences.

Thanks,

Patrick

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