How much micromolar tRNA should I use for crystallisation with a protein?

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Adam B Shapiro

If you know the affinity of the protein-tRNA complex, you can calculate the tRNA concentration needed to saturate binding. The aim is to make sure you have complete occupancy of the protein by tRNA. Also, the tRNA concentration must be at least as high as the protein concentration.

Heide Marie Resch

I agree with Adam. You could also run an analytical size-exclusion chromatography (combined with MALS) after mixing protein and tRNA. This allows you to analyze your sample in more detail, for example you will see if any higher oligomers form or if your sample forms a very homogeneous complex.