Products/Services Used | Details | Operation |
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Gene Synthesis> | All these regulatory elements were assembled using gene synthesis (GenScript) and 40 nucleotides of sequence homology to the insertion sites in the vector backbone (pESC-LEU, Agilent, cat #217452; pESC-TRP, Agilent, cat #217453) were added for gap repair recombination in yeast52,53.... For MAPPIT, half of the cells were stimulated with erythropoietin (2 ng/mL; GenScript) 24 h after transfection. | Get A Quote |
Complementary assays are required to comprehensively map complex biological entities such as genomes, proteomes and interactome networks. However, how various assays can be optimally combined to approach completeness while maintaining high precision often remains unclear. Here, we propose a framework for binary protein-protein interaction (PPI) mapping based on optimally combining assays and/or assay versions to maximize detection of true positive interactions, while avoiding detection of random protein pairs. We have engineered a novel NanoLuc two-hybrid (N2H) system that integrates 12 different versions, differing by protein expression systems and tagging configurations. The resulting union of N2H versions re... More