Plant and Animal Genome Conference 2009 (637k)
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.
"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."
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.