3 replies to this topic

[ston]

In the digest:

pBluescript (one HindIII restriction site)
HindIII

The gel shows 4 bands, I cannot think of why there would be that many. The only thing I can think of is that pBluescript is circlular and the bands are due to different levels of supercoiling, but I feel like if that was the case then the gel would show more of a smear than distinct bands.

Any thoughts?

[phage434]

Have you compared this with uncut plasmid? Incomplete digestion would leave that present and may have (as you suggest) several bands. Perhaps you are trying to cut too much DNA?

[swanny]

Any chance of a gel image?

[T C]

Yeah its uncut DNA