DNA is packaged with histones and non-histone proteins to form chromatin that impinges on all nuclear processes such as transcription, replication and repair. A complete molecular understanding of these processes requires analysis of chromatin context. In this study, we report a simple and robust system using the wheat cell-free protein synthesis to assemble chromatin. We report two assembly protocols, the post-translational nucleosome assembly (PNAP) and co-translational nucleosome assembly (CNAP), both of which resulted in a native and unmodified form of chromatin. In the PNAP, a mixture of four unpurified core histones and chromatin assembly factors was combined with other components and incubated with DNA. In the CNAP, mRNAs encoding for these proteins were incubated with DNA in the translation mixture, and concomitantly assembled chromatin. Furthermore, we developed a rapid one-step method to isolate the assembled chromatin using divalent cations. Isolated chromatin was assessed for the stoichiometry of histones, nucleosome arrays, supercoiling, and as a substrate for histone modification enzymes. Our methods provide a new avenue to assemble customized chromatin to understand chromatin structure and function.