Hey, I was wondering if the following method could work.
-- Say you wanted to make a crude cell extract of E. coli proteins. Like, you want to do DNA ligation very cheaply and effortless. You don't wanna buy ligase.
Let's say you express a huge recombinant ligase or a multimere ligase, and then do a size exclusion centrifugation through a filter.
Basically, you may get a mix of big coli proteins (including the huge ligase) but the nucleases are too small and go right throug the size exlusion filter. Then you add your DNA to the remaining crude protein extract and it will ligate but not degrade the DNA?
Not sure if that's a path worth pursuing, would love your input. There's probably a range of nuclease enzymes out there that come in different sizes
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