Abstract: Deep mutational scanning is a foundational tool for addressing functional consequences of large numbers of mutants, yet a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Provided herein are nicking saturation mutagenesis, a single-day, single-pot saturation mutagenesis method using routinely prepped plasmid dsDNA as input substrate. Reproducibility and convenience of the method are demonstrated through validation by an external research laboratory.

Abstract: Deep mutational scanning is a foundational tool for addressing functional consequences of large numbers of mutants, yet a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Provided herein are nicking saturation mutagenesis, a single-day, single-pot saturation mutagenesis method using routinely prepped plasmid dsDNA as input substrate. Reproducibility and convenience of the method are demonstrated through validation by an external research laboratory.

Abstract: Enzymatically active hydrogenase is synthesized in a cell-free reaction. The hydrogenases are synthesized in a cell-free reaction comprising a cell extract derived from microbial strains expressing at least one hydrogenase accessory protein. In some embodiments, the extracts are produced under anerobic conditions.

Abstract: A folding modular structure is described. The structure typically includes a top panel, which is ultimately the roof deck. Two side panels, a front and a rear panel are connected to the top panel by pivoting mechanisms so that the side, front and rear panels can fold open to create the modular structure. Spacer panels are connected between three of the four total side, front and rear panels. The spacer panels are necessary to provide clearance so that each of the panels can be folded onto the other already folded panels. In the folded configuration, shipping many more modular structures to final destinations is possible.