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E. coli Protein Expression, Purification, and Quantification

skills/ginkgo-cloud-lab/references/ecoli-expression-purification-quantification.md

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E. coli Protein Expression, Purification, and Quantification

URL: https://cloud.ginkgo.bio/protocols/ecoli-expression-purification-quantification Status: Ginkgo Certified Price: $209/sample Turnaround: up to 3 weeks Throughput: Up to 96 constructs in parallel

Overview

Fully automated, end-to-end service that takes DNA constructs from heat shock transformation through bacterial expression, cell lysis, magnetic His-tag (IMAC) bead purification, quantitative yield measurement by A280 absorbance, and LabChip-based purity and size assessment - all in a single unattended workflow. Instruments include the Agilent Bravo 96/384, BioTek MultiFlo, Inheco and Cytomat incubators, a Spark plate reader, and the Revvity LabChip.

Input

  • Protein designs: AA or DNA sequence submitted in CSV format
  • His-tag orientation: N-terminal or C-terminal fusion; linker sequence (default GGGS if unspecified)
  • Known expression notes: Disulfide bonds, cofactor requirements, toxicity concerns, or PTM needs

Output

  • Yield Quantification: A280 per well, converted to protein concentration (mg/mL) via the protein's molar extinction coefficient
  • Expression Confirmation: Fluorescence signal relative to controls
  • Purity & Size Assessment: LabChip-based purity percentage and apparent molecular weight per construct, with virtual gel images
  • OD culture growth data reported per construct
  • Assay Quality Metrics: Per-run quality summary with plate-level controls
  • QC-flagged PDF results report plus downloadable raw CSV files

Automated Workflow

Five phases coordinated across automated instruments:

  1. Transformation with LB recovery (heat shock)
  2. Culture growth and harvest
  3. Cell lysis
  4. Magnetic bead IMAC purification with A280 yield readout (BioTek MultiFlo, Agilent Bravo 96, Bioshake, Spark)
  5. LabChip purity / size assessment (Revvity LabChip)

Ordering

  • Number of Proteins: configurable
  • Number of Replicates: configurable
  • File Upload: CSV, Excel, FASTA, TXT, PDF, ZIP
  • Additional Details: free-text field for special requirements

Use Cases

  • Bacterial expression with purified yield plus purity/size profiling in one run
  • Characterizing up to 96 constructs before scale-up
  • Producing and QC-ing His-tagged protein for downstream work