DNA Molar Pooling Calculator
The Problem: Mass ≠ Molecules
In the laboratory, we are often taught to quantify DNA in nanograms (ng). However, for biological reactions like Next-Generation Sequencing (NGS), Gibson Assembly, or PCR pooling, the weight of the DNA matters much less than the number of molecules.
If you mix a 500bp amplicon and a 2000bp amplicon at the same mass (e.g., 50ng each), you actually have 4 times more molecules of the small fragment. This imbalance can ruin sequencing libraries or lower cloning efficiency.
The Solution
I developed the DNA Molar Pooling Calculator to solve this. It is an R Shiny application that automates the physics of DNA normalization. By calculating based on molarity (fmol) rather than mass, it ensures perfectly balanced pools regardless of fragment size.
Key Features
- Universal: Works for Nanopore, Illumina, PacBio, or standard cloning vectors.
- Batch Mode: You can paste data directly from Excel (Sample Name, Conc1, Conc2…) to process entire plates at once.
- Safety Checks: The app automatically flags samples (in red) that are too dilute to meet your target concentration.
🚀 Live Calculator
You can use the tool directly below, or open it in a new window.
Citation
@online{sánchez-garcía2026,
author = {Sánchez-García, Daniel},
title = {DNA {Molar} {Pooling} {Calculator}},
date = {2026-01-29},
url = {https://danielsangarci.com/posts/dna_molar_pooling_calculator/},
langid = {en}
}