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ecoli-protein-expression-hibit.md

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1# E. coli Protein Expression with HiBiT Quantification
2
3**URL:** https://cloud.ginkgo.bio/protocols/ecoli-protein-expression-hibit
4**Service terms:** https://cloud.ginkgo.bio/terms/ecoli-hibit
5**Reviewed:** 2026-09-30; prices and turnaround below are catalog estimates.
6**Status:** Ginkgo Certified
7**Price:** $79/sample
8**Turnaround:** up to 3 weeks
9**Throughput:** Up to 384 constructs per run
11The catalog says up to 3 weeks, while service terms state 15-20 business days from
12DNA submission. Confirm scheduling in the accepted order.

E. coli Protein Expression with HiBiT Quantification

URL: https://cloud.ginkgo.bio/protocols/ecoli-protein-expression-hibit Service terms: https://cloud.ginkgo.bio/terms/ecoli-hibit Reviewed: 2026-09-30; prices and turnaround below are catalog estimates. Status: Ginkgo Certified Price: $79/sample Turnaround: up to 3 weeks Throughput: Up to 384 constructs per run

The catalog says up to 3 weeks, while service terms state 15-20 business days from DNA submission. Confirm scheduling in the accepted order.

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14## Overview
16Fully 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.

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.

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18## Input
20- **Sequence input:** Protein or DNA in CSV, with HiBiT tag/linker and expression notes. The current page links a CFPS-named `.xlsx` file; verify that its fields cover this E. coli service before uploading.
21- **HiBiT tag orientation:** N-terminal or C-terminal fusion (default GGGS linker if unspecified)

Input

  • Sequence input: Protein or DNA in CSV, with HiBiT tag/linker and expression notes. The current page links a CFPS-named .xlsx file; verify that its fields cover this E. coli service before uploading.
  • HiBiT tag orientation: N-terminal or C-terminal fusion (default GGGS linker if unspecified)
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23## Output
25- **OD600 growth confirmation:** Per-well absorbance readings confirming bacterial growth prior to pelleting
26- **HiBiT luminescence values:** Per-well bcRLU from BMG PHERAstar; preserve the supplied background-correction convention
27- **Normalized expression estimate:** Semi-quantitative expression normalized to an on-plate HiBiT standard curve
28- **QC report:** PDF with per-construct results, process control outcomes, and pass/fail status; raw CSV available

Output

  • OD600 growth confirmation: Per-well absorbance readings confirming bacterial growth prior to pelleting
  • HiBiT luminescence values: Per-well bcRLU from BMG PHERAstar; preserve the supplied background-correction convention
  • Normalized expression estimate: Semi-quantitative 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
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30## Automated Workflow
321. **Transformation** (heat shock)
332. **Autoinduction expression** in lactose-based media
343. **Cell pelleting**
354. **Detergent-based lysis**
365. **HiBiT detection & luminescence read** (BMG PHERAstar)

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)
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38## Ordering
40- **Number of Proteins:** configurable
41- **Number of Replicates:** configurable
42- **File Upload:** CSV, Excel, FASTA, TXT, PDF, ZIP
43- **Additional Details:** free-text field for special requirements

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
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45## Use Cases
47- Very high-throughput expressibility screening in E. coli (up to 384 constructs)
48- Relative expression comparison across large construct sets
49- Early triage before His-tag purification tiers

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
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