Summary

The Research Assistant will design and build GoldenBraid DNA vectors for gene overexpression and CRISPR deletions in the tgrB1–tgrC1 pathway, perform construct validation, and generate stable Dictyostelium lines. They will carry out transformations, clonal isolation, and basic phenotypic analyses by fluorescence microscopy, while maintaining rigorous electronic documentation and producing concise progress summaries for the PI and Assistant Professor. The role follows established SOPs but expects proactive troubleshooting/optimization and staying current with emerging molecular tools.

On‑site laboratory role; schedule may occasionally require evening/weekend attention for time‑sensitive culture passages, selections, or imaging runs.

Job Duties

• Independent design of GoldenBraid vectors (promoter selection, multi‑part assemblies, CRISPR gRNA/repair template design) with PI approval prior to build; execute per SOPs and document rationale, designs, and results in Electronic Lab Notebook.
• Vector construction & validation: scarless assembly (GoldenBraid), colony screening, restriction analysis, and Sanger‑level sequence confirmation; maintain version‑controlled plasmid maps.
• Dictyostelium transformations: select transformants, isolate and propagate pure clones, and prepare quality‑controlled frozen stocks; manage strain records for self‑generated lines.
• Phenotypic characterization: fluorescence imaging on dissecting and compound microscopes; capture and annotate images; advanced quantitative analysis (Fiji/ImageJ, CellProfiler) preferred but not required.
• Troubleshooting & optimization: identify assembly bottlenecks, adjust reaction conditions, and propose protocol updates while preserving compliance with SOPs.
• Data management & reporting: maintain complete electronic lab notebooks; compile weekly summaries and slide/data packets for PI; ensure reproducibility (plasmid/strain linkages, primer logs).
• Collaboration & support: coordinate construct designs and timelines with the PI and lab members; may occasionally assist another technician with general ordering.
• Developmental biology focus: build and test constructs targeting tgrB1–tgrC1 pathway components; integrate cell‑type–specific promoters to drive fluorescent reporters.
• Platform/tooling: use LabArchives and SnapGene for plasmid maps, primers, and gRNA designs; maintain traceability from design → construct → strain → phenotype.
• Strain stewardship: frozen stocks, genotype/phenotype notes, and periodic QC to prevent drift/contamination.
  Safety & compliance: adhere to BSL‑appropriate practices, sterile technique, and BCM policies for data/security and sample handling.
• Innovation expectation: monitor literature/tools (e.g., improved CRISPR systems, cloning strategies) and recommend updates to SOPs for approval.Lab operations boundaries: order and track materials for own projects; general lab ordering handled by another technician (provide ad‑hoc backup only).
• Planning participation (optional): attend relevant design/timeline meetings when helpful to project execution

Minimum Qualifications

• Bachelor's degree in a Basic Science or a related field. 
• Three years of relevant experience.

Preferred Qualifications

• Master's degree in basic science or a related field.
• Hands‑on recombinant DNA work; experience with scarless assembly (GoldenBraid/Golden Gate) strongly preferred; prior Dictyostelium experience desirable; exposure to lambda phage vectors a plus.
• Core skills: sterile technique; eukaryotic cell culture; light/fluorescence microscopy; primer design and PCR; plasmid map management; accurate record‑keeping in ELN.
• Preferred skills: CRISPR gRNA/off‑target analysis; quantitative image analysis (Fiji/ImageJ, CellProfiler); troubleshooting/optimization of cloning and transformations; familiarity with LabArchives, Benchling, or equivalent.