E. coli Protein Expression with HiBiT Quantification

URL: https://cloud.ginkgo.bio/protocols/ecoli-protein-expression-hibit Status: Ginkgo Certified Price: $79/sample Turnaround: up to 3 weeks Throughput: Up to 384 constructs per run

Overview

Fully automated, end-to-end workflow for expressing and quantifying HiBiT-tagged proteins in E. coli. Heat shock transformation is followed by inoculation into lactose-based autoinduction media for target protein expression, then cell pelleting, detergent-based lysis, and HiBiT-based quantification. Results are reported as both raw and standard curve-normalized values, enabling quantitative comparison across up to 384 constructs per run.

Input

• DNA Input: HiBiT-tagged constructs (use the E. coli input template)
• HiBiT tag orientation: N-terminal or C-terminal fusion (default GGGS linker if unspecified)

Output

• OD600 growth confirmation: Per-well absorbance readings confirming bacterial growth prior to pelleting
• HiBiT luminescence values: Raw bcRLU per well from BMG PHERAstar luminescence read
• Normalized expression estimate: Per-construct expression normalized to an on-plate HiBiT standard curve
• QC report: PDF with per-construct results, process control outcomes, and pass/fail status; raw CSV available

Automated Workflow

1. Transformation (heat shock)
2. Autoinduction expression in lactose-based media
3. Cell pelleting
4. Detergent-based lysis
5. HiBiT detection & luminescence read (BMG PHERAstar)

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

• Very high-throughput expressibility screening in E. coli (up to 384 constructs)
• Relative expression comparison across large construct sets
• Early triage before His-tag purification tiers