DNeasy Plant Pro Kits

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

S_8770_DNeasyPlantPro
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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
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Preparations
50
250
DNeasy Plant Pro Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Want to try this solution for the first time?
Get in touch with our team today and request a quote for your DNeasy Plant Pro Kit (50) trial kit.

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

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.

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 table "Selection of plant species processed with DNeasy Kits" and 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”)

Selection of plant species processed with DNeasy Kits
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)

 

See figures

Principle

The DNeasy Plant Pro Kit comprises a novel and proprietary method for fast and easy purification of total cellular DNA from plant cells, tissues and seeds. In addition, this kit can be used to purify bacterial, fungal and viral DNA from plant and root samples. 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.

 

Procedure

 

The DNeasy Plan Pro Kit uses bead-beating technology, whch replaces cumbersome DNA isolation procedures to recover high-quality DNA from the toughest sample types. 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.

 

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 500 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 (1)
Explore our new, easy-to-navigate digital version of the DNeasy Plant Pro Kit Product Profile!
Quick-Start Protocols (1)
Safety Data Sheets (1)
Kit Handbooks (1)
Certificates of Analysis (1)

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

What QIAGEN kit can I use to isolate DNA from food products to test for genetically modified organisms (GMOs)?

For plant-based foods, such as soy, tofu, and cookies, DNA isolation has been successfully carried out using the DNeasy Plant Mini Kit. The QIAamp DNA Stool Mini Kit has been used for isolation of genomic DNA from highly processed foods, and foods that contain high levels of PCR inhibitors, such as chocolate. For meat and processed meats, such as sausage, we suggest the DNeasy Blood & Tissue Kit.

See also the QIAGEN News article "Detection of genetically modified soybean and maize in raw and processed foodstuffs", and the accompanying editorial "Detecting genetically modified organisms in food."

FAQ ID -371
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 h to precipitate the DNA
  • Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15–20 min
  • 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
Does the Epitect Bisulfite Kit also work on DNA extracted from plants?

Bisulfite conversion of unmethylated cytosines into uracils with the EpiTect Bisulfite Kit works on DNA irrespective of the source organism. The DNA template needs to be of high purity for efficient conversion. We recommend to use genomic DNA extracted with our DNA isolation kits for clinical or animal and plant samples as a template for the EpiTect Bisulfite Kit.

FAQ ID -1209
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