By the end of June, the project had moved from broad planning into practical workflow design.
Beta-lactamase selection became the most developed experimental branch. The basic idea was to attach candidate IDP tags to beta-lactamase, expose cells to antibiotic stress, and identify variants that allow better survival. This gave us a potential high-throughput selection system.
However, the details became more complicated. Liquid selection looked attractive because it could screen many cells at once, but we identified a major possible artefact: beta-lactamase released from lysed cells could reduce antibiotic pressure for the whole culture. This would make weak variants look better than they really are. Because of this, plate-based or gradient-based selection remained an important backup.
We also looked more carefully at cloning. Gibson assembly was still attractive, but primer design and possible mispriming against beta-lactamase made it less straightforward than expected. Restriction cloning remained a fallback, although we needed more advice on cost, yield, and timeline.
The computational side also became more disciplined. Instead of relying on one very broad degenerate library, we began considering sequence binning and compression. The aim was to keep libraries within a realistic size while tracking how much sequence information was lost.