Synthetic cell engineers must make their own PURE from scratch to achieve full compositional control of cytosol—for example, to tune energy regeneration systems or try enzyme variants. Typical startup times for getting PURE working currently range from nine months to two years—far longer than almost any cell builder anticipates starting out for such a prevalent technology. This process may involve confronting unexpected challenges in sequence design, protein purification, reagent selection and handling, access to materials, and reproducibility. Our goal with the OnePot Workshop was to reduce as much of this ambiguity as possible and bring startup times down by an order of magnitude.
In May 2025, we wrapped several 
PURE Protocols including 
OnePot PURE workshop module to deliver a workshop in which eight graduate students and postdocs representing three continents built the complete OnePot PURE system from scratch from OpenMTA plasmids in just five days. Workshop participants were able to each build their own energy mix, protein mix, tRNAs, and ribosomes—the four components of PURE—with three out of four components performing at levels comparable to commercial benchmarks (protein mix performed below expectations). They left the workshop with open protocols and OpenMTA plasmids needed to make all four components at their home institutions.
At b.next, we will continue to refine the 
Make OnePot Protein Mix protocol for reliability and this ongoing work will be documented in a series of Developer Notes. Over the next several months, we are continuing to build community and support the participants in making PURE at their home institutions by organizing regular, remote office hours.
“I felt that I got very practical hands on experience that left me feeling confident that I could replicate the protocols in my home lab. I also really loved the opportunity to talk with everyone at b.next and the fellow workshop participants. Having the opportunity to do lab work with others within the field and share advice and hands on tips was incredibly cool and not an experience I have had before.” ”This was such an incredibly cool experience! Getting to do science in such a collaborative environment, with other highly skilled experts was such a unique experience. I absolutely loved all of the conversation that happened throughout the week and cannot think of another environment outside of a workshop like this that would have allowed that to develop.” - Anonymous Workshop Participants
In the meantime, we are sharing the materials that we used to organize and run this workshop with the community in the hopes that it can lower the barrier to working with and customizing the PURE system for use in synthetic cells. The OnePot PURE workshop module includes a schedule and a bill of materials. If you find these materials useful or are interested in organizing your own workshop we would love to hear about it at build@bnext.bio.
This workshop and its materials were developed and organized in close collaboration with Build A Cell, Evan Kalb from the University of Minnesota, Matas Deveikis from Imperial College London, and Charlie Newell from University College London.