Sequential restriction digest

[Nikola Wenta]

Dear all,
I want to do a SacI/SalI digest of my plasmid in order to subclone my
insert (1.2 kb) into another vector. According to NEB, the REs need
different buffers, so I would have to do a sequential digest. Does
anyone have a working protocol for sequential digests? Particularly I
would like to know how to proceed once the first digest i.e. with SacI
has linearized my original plasmid. Would you run a preparative agarose
gel and cut the band out, or would you precipitate the DNA and resuspend
the resulting DNA pellet in the buffer for the subsequent SalI digest?
Thank you for any suggestions!
Niko

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