---
title: "Dose Response (Execution Mode)"
slug: "dose-response-execution-mode"
updated: 2026-09-09T13:57:19Z
published: 2026-09-09T13:57:19Z
canonical: "docs.discngine.com/dose-response-execution-mode"
---

> ## Documentation Index
> Fetch the complete documentation index at: https://docs.discngine.com/llms.txt
> Use this file to discover all available pages before exploring further.

# Dose Response (Execution Mode)

## Scientific Background

To select a practical number of compounds that exhibited high activity in the primary screen for follow-up assays, a cutoff value of the primary %Inhibition was applied. The cutoff value was calculated as the sum of the average percent inhibition of all compounds tested and three times their standard deviation. In this case, 48 unique compounds (1.9% of all compounds screened) were selected as hits.

As before, the number of unique compounds selected for follow-up screening represents a relatively small sample size and therefore were immediately progressed to individual dose response analysis (ED50 determination).

Compounds were "hit-picked" at 10 mM concentration in DMSO and further serially diluted ten times at three-fold dilutions for a total of 10 points per compound. Data was normalized as in the primary assay and curves were plotted and fitted to a four-parameter equation describing a sigmoidal concentration-response curve (e.g., the Hill equation). The reported ED50 values are generated from fitted curves by solving for the x-intercept at the 50% activity level of the Y-intercept. Compounds with ED50 values greater than 0.01 micromolar are considered inactive; compounds with ED50 of equal to or less than 0.01 micromolar are considered active.

![Dose Response Analysis with ASSAY](https://cdn.document360.io/dde9293e-c107-4b3e-9774-9c9c296ed970/Images/Setting/assay-runs-well-validation-3-2.jpg?sv=2026-02-06&spr=https&st=2026-11-10T21%3A30%3A10Z&se=2026-11-10T22%3A30%3A10Z&sr=c&sp=r&sig=tEHiRoMxGCWYJYdfzHBGeRRDGmxb1Gc3Q0NoyqkLi2o%3D)

## Prerequisites

- An existing run template, created using `Design Mode`, following the tutorial in [Dose Response Tutorial](/public/docs/dose-response-design-modes).

Files to use:

- [02-Dose_Response_Assay_p96_Raw_Data](https://cdn.document360.io/dde9293e-c107-4b3e-9774-9c9c296ed970/Images/Documentation/discngine-assay-02-dose-response-assay-p96-raw-data.txt?sv=2026-02-06&spr=https&st=2026-11-10T21%3A30%3A10Z&se=2026-11-10T22%3A30%3A10Z&sr=c&sp=r&sig=tEHiRoMxGCWYJYdfzHBGeRRDGmxb1Gc3Q0NoyqkLi2o%3D)
- [02-Sample_Mapping_Dose_Response_Assay_p96](https://cdn.document360.io/dde9293e-c107-4b3e-9774-9c9c296ed970/Images/Documentation/discngine-assay-02-sample-mapping-dose-response-assay-p96.txt?sv=2026-02-06&spr=https&st=2026-11-10T21%3A30%3A10Z&se=2026-11-10T22%3A30%3A10Z&sr=c&sp=r&sig=tEHiRoMxGCWYJYdfzHBGeRRDGmxb1Gc3Q0NoyqkLi2o%3D)

## Run Creation

- Click the `New Run` button to create a new run.
- Select `Execution Mode` as the mode.
- Enter a `Run Name`.
- Select the appropriate `Run Template` (the one saved in Step 9 of `Design Mode`).
- Go to the `File Reading` step.

## File Reading

- The plate format (`96 (8x12)`) is already set.
- The reader named `Analyst GT (TAB)` is already fixed.
- Upload the file `02-Dose_Response_Assay_p96_Raw_Data`.
- Go to the `Raw Data File` step.

## Raw Data File

- Number of measurements per plate is already mapped: `1`.
- The `Readout Alias` is fixed to `READOUT1` and cannot be modified.
- Go to the `Plate Mapping` step.

## Plate Mapping

The Control Layout named **Dose_Response** is already applied to all plates.

- Go to Well Validation step.

## Well Validation

Visualization template selected in Design Mode is already applied.

You can change this template.

For **QC Validation**, select aberrant wells in the visualization and click on the ![Well Validation step: Invalidation button](https://cdn.document360.io/dde9293e-c107-4b3e-9774-9c9c296ed970/Images/Setting/assay-runs-invalidation-button-2-3-4-5-6.png?sv=2026-02-06&spr=https&st=2026-11-10T21%3A30%3A10Z&se=2026-11-10T22%3A30%3A10Z&sr=c&sp=r&sig=tEHiRoMxGCWYJYdfzHBGeRRDGmxb1Gc3Q0NoyqkLi2o%3D) button. Multiple selection is allowed by holding the Ctrl key.

              
              

Plate_ZSCORE_READOUT1 < -2 or Plate_ZSCORE_READOUT1 > 2.

- Go to Sample Mapping step.

## Sample Mapping

- Click on **Edit Sample Information** button.
- Select filling mode **File** and tick **Sample** and **Dose** for content.
- Choose file 02-Sample_Mapping_Dose_Response_Assay_p96.
- Click on **Apply** button.

Samples and doses are filled in.

- Click on **Continue** button.

Samples and doses are mapped for all plates.

- Click on **Sample Analysis Settings** button.

## Sample Analysis Settings

Groups are automatically created (48 groups). Fitting analysis is already applied.

- Go to Sample Analysis.

## Sample Analysis

The template specified in Design mode is applied.

- For **Hits Selection**, select points (“hits”) in visualization and add them to a list (click on the **Add Hit** button). This selection can be exported in CSV format (export button).
- **Lock** your Analysis (**Lock run** button).

## Publish Data

Depends on the Publisher defined in the workflow template.