App Note
A solution for DNA analysis of plants with minimal sample input using phytoGEM in combination with SolisFAST SolisGreen qPCR mix
With increasing sensitivity and decreasing costs associated with DNA analysis, scientists are pushing the limits of the size of the sample analyzed.
These new limitations have broadened to include applications that involve plants and botanical materials. Examples of such applications include, but are not limited to, plant molecular breeding, identification of Chinese herbal medicines in forensic investigations, and species identification prior to trade of agricultural products. Oftentimes, minimal sample is available for the DNA analysis workflow.
Here we report an optimized extraction protocol to use the phytoGEM® kit chemistry (apart from MicroGEM’s PDQeX platform) for those applications that dictate a lower extraction volume. We show how the use of the phytoGEM chemistry with low abundant samples (for example 2-5 mg of leaf or seed sample) is able to provide large amounts of high-quality DNA, suitable for downstream applications.
Furthermore, the combination of MicroGEM’s temperature-driven extraction chemistry with SolisFAST® SolisGreen® qPCR Mix with UNG allows sample to qPCR results in less than 2 hours.
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PDQeX phytoGEM
Suitable for studying plant genotypes and genetic variation, endophytic plant pathogens, genetic mapping, fingerprinting, microsatellite marker-assisted selection, and for evaluating the authenticity of exported plant varieties, botanical extracts and molecular detection of food adulterant.
Produces double-stranded DNA suitable for PCR and qPCR.
Each kit contains PDQeX prepGEM, Histosolv, enhancer, GREEN+ buffer, and extractor tubes.
For processing larger samples, we recommend the PDQeX phytoGEM HC kits. These kits have the traditional advantages that come with using the PDQeX Nucleic Acid Extractor but also include the new HC extraction cartridges.
These new HC extraction cartridges allow for more sample to be loaded into the cartridge before the purification matrix becomes overloaded. This results in a higher final yield of extracted DNA, meaning the PDQeX phytoGEM HC kits are better suited for sequencing workflows where a higher yield of DNA is encouraged.