We have a recombinant protein (plant virus) that binds RNA. We would like to crystallize this protein, thus we would like it free of nuclei acids. Absorption spectra indicates a significant amount of nucleic acids bound. How can we remove this- we need a protocol that is cheap as the volumes are large?
You can try hydroxyapatite. Relatively cheap and there are plenty of online references with protocols.
Hi Arni,
What is the pI of the protein? I could remove nucleic acids from my samples by Ion Exchange (normally Anion Exchange using HiTrap Q column)! Then the nucleic acids, which are strongly negatively charged, would bind tightly to the column and it would normally elute only in high concentration of salt (say more than 1M salt) whereas the proteins would elute earlier.
It worked very well for me!
Cheers,
Izabella
Hi Byong Lee . DNA is more easily sheared into small nucleotides than proteins. An extruder might work and you can follow the DNA shearing on an agarose gel. Small nucleotides can then be separated from protein using prep dialysis or counter current methods.
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