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DNeasy Plant Pro and Plant Kits

For extraction of total cellular DNA from plant cells and tissues or fungi, or genomic DNA from plant cells, tissues and seeds

Features

  • Superior PCR performance with patented Inhibitor Removal Technology (IRT)
  • Rapid extraction of ready-to-use DNA
  • No organic extraction, no ethanol precipitation
  • Highly efficient lysis and release of DNA from tough plant materials and associated plant pathogens
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DNeasy Plant Pro Kit (50)

Cat. No. / ID: 69204

Tissue Disruption Tubes, MB Spin Columns, Buffers, Collection Tubes (1.5 and 2 ml), for 50 preps
$275.00
Kit
DNeasy Plant Pro Kit
DNeasy Plant Kit
Preparations
50
250
Add to cart
DNeasy Plant Pro and Plant Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

The DNeasy Plant Pro Kit has several innovative features to overcome the challenges of DNA extraction from plant tissue and enable recovery of high-quality DNA from the toughest sample types, including strawberry leaf, grapevine leaf, pine needles and various seed types. Increased yields of plant DNA and plant pathogen DNA combined with superior removal of inhibitors ensure high-performance results in sensitive downstream applications.


Want to try the DNeasy Plant Pro Kit for the first time? Request a trial kit quote.


DNeasy Plant Kits provide fast and easy silica-based DNA isolation from plant samples in spin column format. Typical yields are 3–260 μg of high-quality DNA, depending on the samples used (e.g., wheat, maize, Arabidopsis, tomato, tobacco) and binding capacity of the DNeasy silica membrane. DNeasy Plant Kits also provide silica-based DNA purification in a convenient 96-well plate format with typical yields of 1–15 µg DNA, depending on the plant species. DNA isolation from plant tissue is also automatable on the QIAcube Connect.

Performance

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.

See figures

Procedure

The DNeasy Plant Pro Kit uses bead-beating technology, which replaces cumbersome DNA isolation procedures such as CTAB, phenol or chloroform extraction to recover high-quality DNA from the toughest sample types, including strawberry leaf, grapevine leaf, pine needles and various seed types. The DNeasy Plant Pro Kit uses the second generation of QIAGEN’s patented Inhibitor Removal Technology (IRT) to remove PCR inhibitors, including polysaccharides, polyphenolics and other secondary metabolites from plant extracts during the isolation process. Improved IRT, combined with efficient bead beating and lysis chemistry, results in high yields of inhibitor-free DNA that is ready for immediate use in downstream applications, including PCR, qPCR, and RAPD analysis, RFLP analysis, Southern blotting and next-generation sequencing.


Samples are added to the Tissue Disruption Tube which contains a specially shaped bead and a buffer for rapid homogenization (see figure " Rapid homogenization in Tissue Disruption Tubes"). Cell lysis and DNA release occur by mechanical and chemical methods. Released genomic DNA is cleared of PCR inhibitors using QIAGEN’s second-generation Inhibitor Removal Technology (IRT) and then captured on a silica membrane in a spin column format. DNA is then washed and eluted from the membrane and is ready for PCR, qPCR, NGS and other downstream applications.


An optional Phenolic Separation Solution step helps provide pure DNA even from samples with high polyphenolic compounds, such as pine or grape leaves by breaking the bonds between DNA and phenolics. This step prevents DNA loss that would otherwise occur in these samples.


The DNeasy Plant Pro Kit is used with a vortexer or high-velocity bead beating, based on sample needs. Vortex methods work with soft leaf tissue. High-velocity bead beaters, like the PowerLyzer 24 Homogenizer or the TissueLyser II, break down tougher plant material such as roots, seeds, stems or challenging leaf tissues. Furthermore, purification of DNA using the DNeasy Plant Pro Kit can be fully automated on the QIAcube Connect.


Samples are first mechanically disrupted and then chemically lysed (see flowchart " DNeasy Plant and DNeasy 96 Plant procedures"). RNA is removed by RNase digestion during lysis. Cell debris, precipitated proteins and polysaccharides are removed and the sample is homogenized by centrifugation through a QIAshredder spin column. Buffering conditions are adjusted and the lysate is loaded onto a DNeasy Plant spin column or 96-well plate. During a brief spin, DNA selectively binds to the silica membrane while contaminants pass through. The remaining contaminants and enzyme inhibitors are removed in one or two efficient wash steps. Pure DNA is then eluted in water or low-salt buffer, ready for use.


Purification of DNA using the DNeasy Plant Mini Kit can be automated on the QIAcube Connect.

See figures

Applications

DNeasy Plant Kits provide purification of ready-to-use DNA from plant samples, including plant cells, plant tissues and fungi.
The DNeasy Plant Pro Kit is designed for:

  • DNA extraction from plants and plant-associated microorganisms
  • PCR and NGS analysis
  • Marker-assisted breeding
  • Plant pathogen research
  • Studies on genetically modified plants
  • Detection of resistance traits

Comparison of DNeasy Plant Pro and Plant Kits

Features DNeasy Plant Mini Kit DNeasy Plant Maxi Kit DNAeasy 96 Plant Kit DNeasy Plant Pro Kit
Applications PCR, qPCR, blotting, next-generation sequencing PCR, qPCR, blotting, next-generation sequencing PCR, qPCR, blotting, next-generation sequencing PCR, qPCR, blotting, next-generation sequencing
Elution volume 50–200 µl 500 µl – 2 ml 100–200 µl 50–100 µl
Format Spin column Spin column 96-well plate Spin column
Main sample type Plant samples Plant samples Plant samples Plant samples and seeds
Bead size N.A. N.A. N.A. 5/32” (3.9 mm) ballcone
Binding capacity Up to 50 µg Up to 50 µg Up to 50 µg (per well) Up to 50 µg
Processing Manual Manual Manual Bead beating
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein DNA DNA DNA DNA
Sample amount Up to 100 mg Up to 1 g Up to 50 mg Up to 100 mg
Technology Silica technology Silica technology Silica technology Silica technology
Throughput Varies Varies 96 or 192 samples Varies
Time per run or per prep <1 hour <2 hours <2 hours (192 samples) 45 minutes (24 samples)
Typical yield (from 50 mg starting material) Up to 30 µg Up to 260 µg Up to 15 µg Up to 30 µg

N.A. = Not applicable

Supporting data and figures

Resources

Brochures & Guides (2)
Introducing QIAseq
PDF (450KB)
Accelerate your NGS performance through Sample to Insight solutions
Explore our new, easy-to-navigate digital version of the DNeasy Plant Pro Kit Product Profile!
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Kit Handbooks (2)

Publications

Identification of a gene in the process of being lost from the genus Agrostis.
Li HM; Rotter D; Bonos SA; Meyer WA; Belanger FC;
Plant Physiol; 2005; 138 (4):2386-95 2005 Jul 1 PMID:15995002
Power and limitations of the chloroplast trnL (UAA) intron for plant DNA barcoding.
Taberlet P; Coissac E; Pompanon F; Gielly L; Miquel C; Valentini A; Vermat T; Corthier G; Brochmann C; Willerslev E;
Nucleic Acids Res; 2006; 35 (3):e14 2006 Dec 14 PMID:17169982
Defective RNA processing enhances RNA silencing and influences flowering of Arabidopsis.
Herr AJ; Molnàr A; Jones A; Baulcombe DC;
Proc Natl Acad Sci U S A; 2006; 103 (41):14994-5001 2006 Sep 28 PMID:17008405
Genetic diversity in three groups of barley germplasm assessed by simple sequence repeats.
Matus IA; Hayes PM;
Genome; 2002; 45 (6):1095-106 2002 Dec PMID:12502254
Transposition-based plant transformation.
Yan H; Rommens CM;
Plant Physiol; 2006; 143 (2):570-8 2006 Dec 1 PMID:17142486

FAQ

Are there important considerations for plasma generation and urine handling?

It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.

Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA or citrate as anticoagulant to prevent the release of genomic DNA into the plasma fraction.

Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL-pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.

FAQ ID - 3699
Do you have a protocol for the isolation of DNA from tofu?
Yes, please follow the User-Developed Protocol 'Isolation of DNA from tofu using the DNeasy Plant Mini Kit' (DY09).
FAQ ID -932
How can I precipitate genomic DNA using isopropanol?

Alcohol precipitation is commonly used for concentrating, desalting, and recovering nucleic acids. Since less alcohol is required for isopropanol precipitation, this is the preferred method for precipitation of DNA from large volumes. In addition, isopropanol precipitation can be performed at room temperature, which minimizes co-precipitation of salt that interferes with downstream applications.

 

Procedure

  1. Adjust the salt concentration, for example, with sodium acetate (0.3 M, pH 5.2, final concentration) or ammonium acetate (2.0–2.5 M, final concentration).
  2. Add 0.6–0.7 volumes of room-temperature isopropanol to the DNA solution and mix well.
  3. Centrifuge the sample immediately at 10,000–15,000 x g for 15–30 min at 4°C
  4. Carefully decant the supernatant without disturbing the pellet.
  5. Wash the DNA pellet by adding 1–10 ml (depending on the size of the preparation) of room-temperature 70% ethanol. This removes co-precipitated salt and replaces the isopropanol with the more volatile ethanol, making the DNA easier to redissolve.
  6. Centrifuge at 10,000–15,000 x g for 5–15 min at 4°C.
  7. Carefully decant the supernatant without disturbing the pellet.
  8. Air-dry the pellet for 5–20 min (depending on the size of the pellet).
  9. Redissolve the DNA in a suitable buffer.

Tip: Use a buffer with a pH of 7.5–8.0, as DNA does not dissolve easily in acidic buffers. Often distilled water can have an acidic pH. The addition of EDTA protects the DNA from DNase digestion.

Tip: High-molecular-weight DNA, such as genomic DNA, should be redissolved very gently to avoid shearing. If the DNA pellet does not dissolve easily, heat at 55°C for 1–2 h with gentle shaking.

FAQ ID -2953
How do I perform a DNA precipitation to concentrate my sample?
  • Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample
  • Mix, and store at -20°C for at least 1 hour to precipitate the DNA
  • Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15-20 minutes
  • Pour off the ethanol and wash the pellet twice with room-temperature 70% ethanol
  • Allow the DNA pellet to air-dry
  • resuspend the DNA in a suitable volume of sterile TE buffer or distilled water
FAQ ID -305
How should I store plant material for DNA isolation using the DNeasy Plant Kit?

If storage is required, we recommend to freeze freshly harvested plant material immediately in liquid nitrogen and store it at –70°C for later use with the DNeasy Plant Kits. Alternatively, dry the plant material in the presence of a 10-fold excess of silica gel and seal it in a plastic bag (if the material is not dried completely after 12 hours the DNA is likely to degrade. See Mark W. Chase, Harold H. Hills; Taxon, Vol. 40, No. 2,  1991, pp. 215-220: 'Silica Gel: An Ideal Material for Field Preservation of Leaf Samples for DNA Studies').

For more information on sample storage, see also QIAGEN News article 2004 e8 'General considerations for the storage of sample material prior to DNA purification'.

To avoid overloading the DNeasy Mini Spin Column do not use more than 100 mg fresh or frozen tissue, or 20 mg dried plant tissue, for each DNA preparation.

 

FAQ ID -114
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728
Can the QIAshredder Maxi Spin Columns from the DNeasy Plant Maxi Kit be used for RNeasy samples?

The QIAshredder Maxi spin columns  from the DNeasy Plant Maxi Kit are not recommended for use in conjunction with QIAGEN's RNA isolation products. The filter can only tolerate low centrifugal forces not sufficient to shear genomic DNA. If genomic DNA is not sheared, there will be an increase in genomic DNA contamination in the purified RNA.

For an alternative to QIAshredder homogenization in combination with RNeasy Midi/Maxi Preps, please see FAQ 560.

FAQ ID -616
What is the composition of buffer AE?

The composition of Buffer AE is:

  • 10 mM Tris-Cl
  • 0.5 mM EDTA; pH 9.0.
FAQ ID -730
How do I safely inactivate biohazardous flow-through material?

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12
Do I need to have EDTA in the buffer in which I am going to store my isolated genomic DNA?

EDTA chelates divalent cations which are required for nuclease activity. While the genomic DNA (gDNA) extracted using QIAGEN products, should not have any nuclease activity, it is possible to introduce nucleases during repeated long-term access of the DNA. EDTA helps to prevent any nuclease activity introduced after the genomic DNA extraction procedures.

However, if the gDNA is stored frozen at -20oC or -80oC, nuclease activity is much reduced. It may be possible to leave EDTA out of the storage buffer without negative consequences when samples are kept under these conditions, and when repeated freeze-thaw cycles are avoided.

We do recommend however that gDNA be stored in a neutral to a slightly basic buffered solution (e.g. 10 mM Tris-Cl pH 8.5 to 9.0) to prevent DNA degradation by acid hydrolysis. Note that deionized water mostly has an acidic pH.

FAQ ID -754