A Modular High-Throughput Pipeline for Automated Microbial Strain Engineering: From Colony Picking to Phenotypic and Genotypic Analysis v2

Résumé fourni par la source

Microbial strain engineering has evolved to incorporate guide RNA, oligo pools, and random insertion libraries with millions of combinations to decipher mechanisms of bioproduct production in host organisms. However, effective high-throughput screening of individual colonies for both genotypic and phenotypic traits remains challenging. We present an integrated high-throughput automation pipeline capable of handling over 5,000 samples daily for comprehensive strain engineering tasks. This system bridges the gap from initial colony selection through genotyping and phenotypic analysis using multiple analytical platforms. Our modular workflow consists of several integrated processes: High-throughput colony pickingusing automated colony picking systems Media preparation and distributionwith customizable volumes and plate formats Glycerol stock and production plate preparationwith flexible options for culture transfer, replication, and preservation Culture transfer for scaled productionallowing for additional stamping operations with customizable media compositions Sample preparation for genotyping and phenotypingfor specialized assays (proteomics, RapidFire) and NGS phenotyping with configurable transfer parameters This pipeline offers significant flexibility, allowing researchers to employ individual modules independently or reconfigure workflows according to specific experimental requirements. The system's standardization enhances reproducibility while accelerating strain engineering cycles for synthetic biology and bioproduction applications.

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