Has anyone performed a polysome profile of in vitro translation in PURExpress (NEB) system?

17 October 2018 0 4K Report

When I perform in vitro translation in NEB's PURExpress system, using DNA as a template, and then loaded the reaction on to a 10-40% sucrose gradient. However, most of reaction contains free subunits, and only a tiny fraction (~10%) exists as polysomes.

There is no literature available on polysome profiles from PURExpress systems. I understand that PURExpress reactions are very dilute in comparison to in vivo conditions. Any other suggestions on why this occurs will be most valuable.

Please also let me know if you have observed good polysomes in this system.

Thanks!

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Science method

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