The DNeasy Plant Pro Kit – the newest member of the trusted DNeasy Plant family – enables purification of significantly higher yields of DNA from the toughest sample types, including strawberry leaf, grapevine leaf, pine needles and various seed types (see figure “ Significantly higher yields of pure DNA”). Patented Inhibitor Removal Technology (IRT) provides superior removal of inhibitors without using harsh chemicals, yielding pure DNA with less PCR inhibition (see figure “ Efficient removal of PCR inhibitors”).
When CT values of PCR reactions with plant DNA eluates containing possible inhibitors were compared to CT values of the PCR reaction with water added as control which does not inhibit amplification of the IC DNA, the eluate from the DNeasy Plant Pro Kit showed no inhibition.
With the DNeasy Plant Pro Kit, yields of DNA purified from strawberry leaf – a particularly tough plant sample – were significantly higher than those obtained using a kit from Supplier T and Supplier Z. Furthermore, the DNeasy Plant Pro Kit provided superior removal of inhibitors and the DNA eluate showed no inhibition (see figure “ Significantly higher yields and superior inhibitor removal”). The DNeasy Plant Pro Kit can also purify bacterial, fungal and viral DNA from plant and root samples.
The kit can be successfully combined into a workflow with proven products optimized for next-generation sequencing (NGS) (see figure “ Optimized NGS workflow”). Rapid and reliable identification of plant pathogens is crucial to avoid disease spread and facilitate effective disease management. Plant and pathogen DNA co-purified using the DNeasy Plant Pro Kit enables successful identification of a range of pathogens (see figure “ Successful identification of plant pathogens by NGS”).
DNeasy Plant Kits allow rapid and efficient isolation of high-quality DNA from a wide variety of plant species and tissue types, including the most demanding sources (see table "Selection of plant species processed with DNeasy Kits"). Samples may be fresh, frozen or dried. The optimized DNeasy Plant procedure incorporates the QIAshredder spin column, a unique filtration and homogenization column that efficiently removes cell debris and improves sample handling following lysis.
The typical yield is 3–260 µg, with a sample size of up to 1 g wet weight, and an elution volume of 50 µl to 2 ml. DNA yields vary between different species and tissues depending on genome size, ploidy, cell number and tissue sample age.
|Abies alba (silver fir) ||Nicotiana tabacum (tobacco) |
|Aesculus hippocastanum (horse chestnut) ||Oryza sativa (rice)4 |
|Arabidopsis thaliana (thale cress) ||Pelargonium sp. (geranium)4 |
|Avena sp. (oat) ||Petunia sp.4 |
|Brassica napus (oilseed rape) ||Pinus sylvestris (Scotch pine), P. brutia5 |
|Brassica oleracea (kohlrabi) ||Populus tremula (aspen) |
|Chicorium endivia (chicory) ||Pseudotsuga menziesii (Douglas fir) |
|Citrullini lanatus (water melon) ||Quercus robur, Q. petrea (oak)6,7 |
|Egeria sp. ||Rhododendron sp.2,4 |
|Fagus sylvatica (beech)1 ||Rubus idaeus (raspberry) |
|Helianthus spp. (sunflower) ||Solanum tuberosum (potato) |
|Hordeum vulgare (barley)2 ||Sphagnum palustre (moss) |
|Humulus sp. (hops) ||Spinacia oleracea (spinach) |
|Hydrilla sp. ||Taxus baccata (yew) |
|Kalanchoe spp. ||Triticum aestivum (wheat)4 |
|Lupinus sp. ||Ulmus glabra (elm)6 |
|Lycopersicon esculentum (tomato)3 ||Vitis spp. (grape)6 |
|Myriophyllum sp. ||Zea mays (maize) |
The DNeasy Plant procedure yields pure nucleic acid, free of polysaccharides and other secondary metabolites often copurified using conventional methods. Such impurities can interfere with spectrophotometric readings and inhibit enzymatic reactions. Absorbance scans of DNeasy purified DNA show a symmetrical peak at 260 nm (see figures " DNA purity from oak leaves and pine needles"), confirming that the DNA is free of impurities, including enzyme inhibitors. DNeasy purified DNA is sized up to 40 kb (see figure " Pure DNA (20–25 kb) for restriction analysis"). Purified DNA can be used in a wide range of applications (see figures " PCR analysis" and " RAPD analysis").
High-quality DNA can be isolated from 96 or 192 samples of plant leaf tissue in less than 2 hours. The simple 96-well procedure provides highly reproducible yields of total cellular DNA (see figure " Uniformity of DNA yields from 96 wheat samples"). The typical yield is 1–15 µg per 50 mg plant leaf tissue, with an elution volume of 100–200 µl. DNA yields vary between species depending on genome size, ploidy, cell number and tissue sample age.