Job Duties and Responsibilities:
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Molecular cloning and construct generation – 25%
Design and build plasmids for engineered T7 polymerases, optogenetic domains, protein binders, reporters, and assay controls using PCR, Gibson/Golden Gate assembly, mutagenesis, and sequence verification.
• Protein expression, purification, and QC – 20%
Express and purify polymerases, binders, and optogenetic protein components (primarily from bacteria) using affinity chromatography/FPLC and standard QC methods such as SDS-PAGE, concentration assessment, and storage optimization.
• In vitro functional assay development and execution – 25%
Develop and run cell-free transcription and polymerase assays, including time-course transcription assays, one- or few-base incorporation assays, and assays using modified nucleotides such as fluorescently labeled or biotinylated bases.
• Optogenetic characterization and readout integration – 15%
Perform light-control experiments, fluorescence/plate-reader assays, and other optical measurements needed to evaluate gating behavior, kinetics, and retained function of engineered constructs.
• Sequencing sample preparation and data/QC handoff – 10%
Prepare samples for sequencing-based readouts, organize construct/sample metadata, and maintain clean experimental records that support downstream analysis and sponsor reporting.
• Project support, documentation, and collaboration – 5%
Maintain SOPs, reagent inventories, and experimental documentation; prepare figures/tables for meetings and reports; and coordinate day-to-day bench workflows with the PI, trainees, and collaborating groups.