DNeasy Plant Kits

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

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DNeasy Plant Mini Kit (50)

Cat. No. / ID: 69104

50 DNeasy Mini Spin Columns, 50 QIAshredder Mini Spin Columns, RNase A, Buffers, Collection Tubes (2 ml)

Column typePlate type

Mini

Maxi

96 well

Preparations

250

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✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Pure DNA, free from contaminants and enzyme inhibitors
Rapid extraction of ready-to-use DNA
No organic extraction, no ethanol precipitation

Product Details

The 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. The 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 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"). The 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.

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)

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.

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"). The purified DNA can be used in a wide range of applications (see figures " PCR analysis" and " RAPD analysis").

See figures

DNA purity from oak leaves and pine needles.

Pure DNA (20–25 kb) for restriction analysis.

PCR analysis.

RAPD analysis.

Principle

The DNeasy Plant Kits use advanced silica-membrane technology and simple spin procedures to isolate highly pure total cellular DNA from plant tissues and cells or fungi. The DNeasy technology replaces cumbersome DNA isolation procedures such as CTAB, phenol or chloroform extraction. Alcohol precipitation is also not necessary, since the purified DNA is ready for immediate use.

Procedure

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 the DNeasy Plant Mini spin column. During a brief spin, DNA selectively binds to the silica membrane while contaminants pass through. 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.

See figures

DNeasy Plant and DNeasy 96 Plant procedures.

Applications

DNeasy Plant Kits provide purification of ready-to-use DNA from plant samples, including plant cells, plant tissues and fungi.
The DNeasy Plant 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

DNA purity from oak leaves and pine needles.

Spectrophotometric scans (220–320 nm) of DNA isolated from pine needles using the method of Dellaporta, CTAB, or the DNeasy Plant Mini Kit. Pure DNA typically shows a symmetrical peak at 260 nm and a smooth profile. Polysaccharides and other secondary metabolites, often copurified with plant DNA isolated using traditional methods, can interfere with OD readings (A260/ A280), leading to errors in determination of concentration and purity.

Resources

Accelerate your NGS performance through Sample to Insight solutions

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

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

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

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

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).
Add 0.6–0.7 volumes of room-temperature isopropanol to the DNA solution and mix well.
Centrifuge the sample immediately at 10,000–15,000 x g for 15–30 min at 4°C
Carefully decant the supernatant without disturbing the pellet.
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.
Centrifuge at 10,000–15,000 x g for 5–15 min at 4°C.
Carefully decant the supernatant without disturbing the pellet.
Air-dry the pellet for 5–20 min (depending on the size of the pellet).
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

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

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

The composition of Buffer AE is:

10 mM Tris-Cl
0.5 mM EDTA; pH 9.0.

FAQ ID -730

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

Yes, please follow the User-Developed Protocol 'Isolation of DNA from tofu using the DNeasy Plant Mini Kit' (DY09).

FAQ ID -932

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

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