Journal: bioRxiv

Article Title: Large-scale exploration of protein space by automated NMR

doi: 10.64898/2026.02.16.706194

Figure Lengend Snippet: Design pipeline (top); de novo backbones were generated using either RFdiffusion, or Proteína, followed by sequence design with ProteinMPNN and filtering with Boltz-1. Experimental validation (bottom); designs were ordered as synthetic genes, cloned into expression vectors, and produced in E. coli from 15 N-labelled autoinduction media in 96-deepwell plates (left). Samples were purified by immobilized metal affinity chromatography in 96-well plate format, followed by high-throughput size-exclusion chromatography (middle). NMR fingerprints are obtained in 45 min for all produced samples.

Article Snippet: Assembled plasmids were transformed into lab-made chemically competent BL21(DE3) E. coli cells (NEB #C2527) obtained with an adapted protocol from Yang et al. To each 1 μL Golden Gate reaction, 8 μL of ice-cold KCM (500 mM KCl, 250 mM MgCl 2 , 150 mM CaCl 2 ) and 8 μL of ice-cold competent cells were added, followed by incubation on ice for 30 minutes.

Techniques: Generated, Sequencing, Biomarker Discovery, Clone Assay, Expressing, Produced, Purification, Affinity Chromatography, High Throughput Screening Assay, Size-exclusion Chromatography

Journal: Bio-protocol

Article Title: A One-Step Method for Efficient Purification of Functional Cas9 Protein

doi: 10.21769/BioProtoc.5594

Figure Lengend Snippet: SDS-PAGE analysis of the purification of Ub-Cas9. (A) Early-elution fractions. Lane 1, E. coli lysate; 2, flowthrough; 3, wash solution; E1–E3, eluted fractions with 80 mM imidazole; E4–E6, eluted fractions with 500 mM imidazole. (B) Late-elution fractions with 500 mM imidazole (E7–E15). For both panels, lanes B1–B5 contain 1–5 μg of BSA (asterisks) as a quantitative standard. The Ub-Cas9 protein is indicated by red arrowheads. M, protein standards.

Article Snippet: E. coli BL21 (DE3) cells (NEB, catalog number: C2527H) 5.

Techniques: SDS Page, Purification