NGS Equimolar Pooling Calculator

A web-based tool for calculating equimolar pooling volumes from complex sub-pools for Next-Generation Sequencing (NGS).

About

This calculator helps researchers determine the correct volumes to take from each pool when combining multiple complex sub-pools of NGS libraries for sequencing. It ensures that each individual sample contributes an equal number of molecules to the final sequencing run, resulting in more balanced coverage.

Features

• Calculate equimolar pooling volumes for any number of input pools
• Support for both calculated (estimated / target) and measured concentrations
• Option to maximize the usage of original pools
• Adjustable final volume
• Real-time results with detailed metrics
• Verification of equimolarity across samples

Live Calculator

The calculator is available at: https://bwprice.github.io/ngs-equimolar-pooling-calculator/

Usage

1. Enter information for each input pool:

   • Molarity that each sample was normalized to in the sub-pool - i.e. the target molarity when pooling (nM)
   • Total pool volume (μl)
   • Number of samples in the pool
   • (Optional) Measured concentration of each sub-pool from lab quantification

2. Choose whether to:

   • Use measured concentrations (if available from Qubit, Bioanalyzer, qPCR, etc.)
   • Maximize usage of original pools
   • Set a specific final pool volume

3. The calculator will automatically determine:

   • How much volume to take from each sub-pool
   • The percentage of each sub-pool that will be used
   • The final nM contribution per sample
   • Total samples in the final pool

Installation for Local Development

1. Clone this repository:

   git clone https://github.com/bwprice/ngs-equimolar-pooling-calculator.git
   cd ngs-equimolar-pooling-calculator
   

2. Install dependencies:

   npm install
   

3. Start the development server:

   npm start
   

4. Open http://localhost:3000 to view it in your browser.

Deployment

To deploy to GitHub Pages:

npm run deploy

Contributing

Contributions are welcome! Please feel free to submit a Pull Request.

License

This project is licensed under the MIT License - see the LICENSE file for details.

Citation

If you use this tool in your research, please cite:

Ben Price, NGS Equimolar Pooling Calculator, GitHub Repository, 2025, 
Available at: https://github.com/bwprice/ngs-equimolar-pooling-calculator