Extracting DNA from plants can be challenging because of their tough cell walls and the inhibitory polysaccharides and tannins released during the extraction process. Currently available methods address these difficulties by using tissue grinding followed by silica beads, columns or organic solvents. These procedures are multistep making them labour intensive and time-consuming.
The phytoGEM extraction system is a simple closed-tube method for extracting PCR-ready plant DNA. It uses a collection tool that crushes the material onto storages cards. Punches from the card are lysed using an aggressive thermophilic proteinase combined with a cocktail of cell wall degrading glycosyl hydrolases. The extraction is performed automatically in the ZyGEM PDQeX2400 device which both lyses the cells and purifies the DNA through a proprietary polymer matrix.
All-up, it takes around 15 minutes to go from leaf to DNA … And most of that time is hands-off.
To demonstrate the extraction efficiency, plant DNA extraction system, six plants were selected: Arabidopsis thaliana, Oryza sativa (Rice), Zea mays (Maize), Triticum aestivum (Wheat), Solanum lycopersicum (Tomato) and Citrus limon (Lemon). The results from qPCRs and end-point PCRs are shown below.
qPCR plots of alcohol dehydrogenase (adh) gene fragments amplified from phytoGEM extractions from 6 plants.
Agarose gel images showing amplicons generated from plant extractions. Fragments were generated using primer combinations targeting sequences in 5 genes: adh (alcohol dehydrogenase), act2 (actin 2), matk (maturase K), rbcL (RuBisCO large subunit) and psbA (photosystem II) ranging from ~200–2300 bp.
- (A) Arabidopis: 1-4: act2, 5-6: matK, 7-8: rbcL, 9-10 psbA, 11-12: adh
- (B) Rice: 1-2: rbcL, 3-4: psb, 5-14: various fragments of adh.
- (C) 1-2: lemon adh, 3-4: lemon rbcL, 5: lemon psb, 6-7: maize adh, 8-9: maize psb, 10-11: maize rbcL, 12-13: tomato psb, 14-15: tomato rbcL.