# 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](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 ```