Grant: Engage Grants Program

Environmental science relies more than ever before on DNA based evidence acquired from samples of varied sources (ecology, population genetics, metagenomics, pathology, etc). While samples acquired from water, plant or insect can yield nice quality DNA, extractions performed from soil are hazardous. Soil composition is highly heterogenous and many substances contained in soil can affect DNA yield, purity and subsequent downstream reactions. To date, soil sample is rarely use for specie specific DNA quantitation since DNA amplification is heavily affected by inhibitors from soil samples. Although several protocols have been devised to minimize the effect of inhibitor, none is universal to all soil type and all have their pitfalls. Our laboratory uses soil samples from black spruce, white spruce, and pines (several species) stands. Once DNA is extracted from these samples amplification is very challenging due to a high concentration of humic acid and other organic acids, although a presence/absence test can sometime be performed, quantitation from such samples is not possible. Bio-Rad Laboratories is a world leader in consumable and equipment related to DNA amplification and quantitation. Bio-Rad has launch droplet digital PCR (ddPCR) during which a PCR reaction is fragmented in several thousands miniPCR reactions and quantified separately, according to Bio-Rad this technology is much less prone to inhibitor induced bias than traditional PCR or real-time PCR. Since BioRad wants to increase their market share in DNA-based environmental samples, demonstrating that ddPCR can outperform current available technologies would give them an edge over its competitors in this field. Therefore, Bio-Rad want to compare state-of-the-art real-time PCR over ddPCR with our heavily contaminated soil samples to seize additional market shares in the field of environmental DNA quatitation.