Direct DNA Amplification from Plants

Genetic marker-assisted breeding of plants is limited by the need to purify nucleic acids from plant tissues.  Non-commercial protocols for preparing crude extracts suffer from limited utility across diverse plant species, limited utility in high-throughput applications, and the use of high temperature acids and bases. 

We have developed a reagent for preparing crude DNA extracts that are compatible with endpoint PCR, dye-based real time PCR, and probe-based real time PCR in a variety of plant species.  Lysis can be performed in 5 minutes, uses no toxic or reactive chemicals, and is compatible with high-throughput automation.  Using single 6-8mm leaf punches from tomato, soy, corn, and cucumber leaf, lysates prepared with the Promega reagent showed earlier and more consistent amplification than those prepared using Hot Sodium Hydroxide and Tris (HotSHOT) methods.  Additional plant species, including oil palm, and seeds have also been successfully tested.  Lysates are compatible with standard and fast cycling using GoTaq® qPCR Master Mix and GoTaq® Probe qPCR Master Mix, and can be decanted and stored at 4°C for at least a year, or further purified as needed. 

Direct amplification without purification is now a workflow option for a wider selection of plants, enabling high throughput genetic mapping, genotyping, transgenic screening, and sexing.