Restriction enzyme SBFI failed to cut plasmid - (Sep/04/2005 )

Does anybody use the SbfI from biolabs?

I digest my plasmid which contains SbfI recongnition site but fail to digest my plasmid.

The buffer is the optimal one NEBuffer4, at 37c.

Any suggestion?

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There's probably nothing wrong with the restriction enzyme.

It's more likely that the functional map for your plasmid is wrong and there is no sbfI site.

How are you checking it for cutting, btw?

-Matt

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As Matt has asked, how did you know that your plasmid didn't cut? Have you run linearized plasmid side by side with uncut plasmid?

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Yes, I have run linearized plasmid by uncut plasmid, the bans are nearly the same. Both have three bands. Does it show that the plasmid failed to be cut?

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