Hello,
Me and my fellow undergrads are attempting to locate RSVGN insert to pali2 vector. The problem earlier was, we ran out of Qiagen MinElute kit used for clean up after restriction digest. I ended up calling Norgen Biotek to ask if we can use their pcr purification kit to clean up digest and they said it should be fine.. so we used it.
Final products have concentrations of 10ng/ul, 10.3 ng/ul, and 20 ng/ul. And an a 260/280 of 1.74, 1.56, and 1.66 respectively.
My question is, since the DNA is not "pure" is this gonna affect our ligation?
Should we redo the process before we attempt to clone ?
Thank youu.
Hi,
I have ligated several preparations from PCR and especially from agarose gel cleanup kits in that 260/280 range sucessfully.
With our nanodrop-like photometer lower DNA samples (approx. < 30 ng/µl) generally have a lower 260/280 ratio, (1.6 to 1.8) even if they were diluted from a more concentrated sample with a perfect ratio of 1.8, which may be explained by measurement variation from the instrument at low sample concentrations. If your preps are not especially valuable and may be repeated readily, I would definitely give it a try.
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