Poster from Plant and Animal Genome Conference 2009 (637k)

Applications

·         High-throughput isolation of DNA from ground seed samples for PCR-based analysis, e.g., GMO testing.

The QuickExtract™ Seed DNA Extraction Solution can be used to rapidly and efficiently extract PCR-ready genomic DNA from most ground seed samples using a simple, one-tube protocol that takes only 8 minutes (Fig. 1). The method allows for the inexpensive processing of one to hundreds of samples simultaneously, without centrifugation, spin columns or use of any toxic organic solvent

The procedure is fully compatible with robotic automation, provides a PCR-ready sample, and is reproducible (Fig. 2). Simply add the QuickExtract Seed solution to the ground sample and perform two sequential heating steps. A small aliquot of the sample mix is then used as a template for PCR or qPCR. EPICENTRE recommends PCR optimization with the FailSafe™ PCR PreMix Selection Kit or a fast PCR with the TAQXpedite PCR System. Seeds tested include apple, barley, cotton, maize, oat, rice, rye, sunflower, switchgrass, tomato, and wheat.

Note: Most ground seed material under 10 mg is acceptable for DNA extraction. Extracted DNA from larger sample sizes will not produce satisfactory PCR results.

Benefits

·         Nontoxic reagents, inexpensive processing.

·         Short procedure (8 minutes for the average sample).

·         No centrifugation or spin columns to reduce yields.

·         Suitable for high-throughput or automated workflows.

"Your QuickExtract Seed and PlantAmp helped me resurrect some DNA templates that I was on the verge of abandoning due to lack of quality DNA and subsequent PCR amplification for downstream sequencing."

Millie Burrell
Texas A&M University

Figure 1 (click to enlarge). Overview of the QuickExtract™ Seed DNA extraction procedure.

Figure 2. PCR of DNA extracted from brown rice seeds. Samples (10 mg each) of five different brown rice seeds were used for DNA extraction, and a 1-µl aliquot of each was used for PCR of the single-copy RIA gene. Lane M, 100-bp ladder; lane 1, no-template control; lanes 2-6, PCR products from five different seed samples.