04 November 2019 4 3K Report

I'm trying to linearize large amounts of plasmid (500 µg - 1000 µg). I can easily digest this amount of plasmid, but when I precipitate with ethanol and resuspend in TE, I'm left with a milky white solution. Initially I thought this was due to excess salt precipitation, but I can wash with 70% ethanol until the cows come home and it doesn't get any better, so clearly it's not salt.

I can purify with a column, but the losses are unacceptably high.

After butting my head against the wall for several days, I realized that my problem must be BSA from the restriction digest buffer. If I digest 1000 µg plasmid DNA in NEB cutsmart buffer, there's 5 mg BSA in the reaction. How can I remove the BSA so I'm not left with filthy dirty DNA when I precipitate? Will precipitating the BSA with ammonium acetate work?

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