What?
PURE is a defined set of 108 molecules (proteins, RNAs, small organic molecules, and salts) that recapitulate transcription and translation, and is widely used in synthetic cell engineering. PURE was originally developed in 2001 and is available commercially under the product names PUREfrex (GeneFrontier Corporation) and PURExpress (NEB) at high cost and under proprietary formulation.
Several protocols have since been published to make PURE production more accessible, including TraMOS PURE, PURE 3.0, and OnePot PURE. These protocols have generated significant interest but have proven surprisingly difficult to boot up. Reproducibility is limited by high biological variance intrinsic to existing protocols and DNA distributions as well as a lack of openly available and shareable DNA. Nucleus PURE is a clean sheet open source approach to make PURE construction easy, accessible, and reproducible.
How?
- Grow E. coli expressing proteins
- Lyse E. coli
- Purify proteins
- Exchange buffer and concentrate proteins
- Assemble and run cytosol
PURE Protocols
Nucleus PURE protocols are still in development. We’re still working on making these protocols work robustly & with our newly designed open-source .
In the meantime, feel free to use these protocols. We cannot guarantee that they’ll work for you (yet) but we’re happy to share what we’ve learned here and answer any questions that you may have. Join our mailing list or Contact us! to know when our DNA collection is available and when these protocols are up and running.