Plants have evolved a variety of defense mechanisms against predators and environmental challenges. Among their protective devices are a myriad of compounds, some very complex in structure, that allow plants to repel pests. These protective compounds include polyphenols and tannins sometimes found in seeds and seed coats. The presence of storage carbohydrates and polyphenols can interfere with successful amplification of DNA prepared from seeds. Until now, cumbersome preparation steps were needed to purify analytical amounts of seed DNA.
Epicentre Biotechnologies’ new QuickExtract™ Seed DNA Extraction Solution facilitates the extraction of PCR-ready DNA from a wide variety of seeds, both monocot and dicot. The kit eliminates the need for organic solvents or detergents such as cetyltrimethyl-ammonium bromide that are commonly required for PCR-ready DNA isolation from seeds.
Using the QuickExtract Seed Solution, PCR-ready DNA can be isolated from 10 mg or less of seed material in under 10 minutes. The requirement for germinated seed-leaf DNA can often delay the sale or export of seed. QuickExtract saves the time and effort needed to plant and sprout seeds for PCR-based analysis.
The PlantAmp™ PCR System is specially formulated for PCR from plant DNA that contains polyphenols and other PCR inhibitors. In addition, the system ensures reliable PCR of templates with up to 80% GC content. The combination of these two kits provides rapid DNA extraction and successful PCR from challenging plant seed samples.
Methods and Results
DNA was extracted from seed samples (ground, chipped, or fragmented) using the QuickExtract Seed Solution. The procedure consists of adding QuickExtract Seed Solution to the seed sample, followed by two incubation steps at 65°C and 98°C. PCR was performed using the PlantAmp PCR System with 35 cycles of amplification. PCR products were separated and visualized in 1% or 2% agarose gels. Primer sequences are available upon request from Epicentre.