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Plasmid DNA Preparation> | … Plasmids The plasmid pJK_proJK1_LGK was created by inserting a codon-optimized gene encoding levoglucosan kinase (LGK) (Genscript, Piscataway, NJ) with LEHHHHHH as 95 the C-terminal tag into a pJK-series plasmid [20] using flanking NdeI/XhoI restriction sites … | Get A Quote |
Comprehensive sequence-function mapping involves detailing the fitness contribution of every possible single mutation to a gene by comparing the abundance of each library variant before and after selection for the phenotype of interest. Deep sequencing of library DNA allows frequency reconstruction for tens of thousands of variants in a single experiment, yet short read lengths of current sequencers makes it challenging to probe genes encoding full-length proteins. Here we extend the scope of sequence-function maps to entire protein sequences with a modular, universal sequence tiling method. We demonstrate the approach with both growth-based selections and FACS screening, offer parameters and best practic... More