For purification of total RNA from plants and fungi
- High-quality total RNA in 30 minutes
- No phenol/chloroform extraction
- No CsCl gradients, no LiCl or ethanol precipitation
- Excellent recovery of RNA
- Ready-to-use RNA for any downstream application
The RNeasy Plant Mini Kit includes QIAshredder spin columns for homogenizing and filtering viscous plant or fungal lysates, and RNeasy spin columns for purifying up to 100 μg of high-quality RNA using silica-membrane technology. Purification can be fully automated on the QIAcube Connect. The kit can also be used in combination with a TissueRuptor or TissueLyser system, which provide efficient disruption and homogenization of plant samples.
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製品名
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RNeasy Plant Mini Kit (50)
50 RNeasy Mini Spin Columns, 50 QIAshredder Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
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74904
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RNeasy Plant Mini Kit は分子生物学実験用です。疾病の診断、治療または予防の目的には使用することはできません。
RNeasy Plant Mini procedure.|Detection of viruses.|Tissue specificity of histone H4 expression.|
|Total RNA was isolated from budwood obtained from infected apple (1), cherry (2), pear (3), and grapevine (4–8), leaf tissue from black currant (9, 10), and dormant tuber tissue from infected potato (11, 12) using a modification of the RNeasy protocol. Viral RNA was amplified using a one-tube RT-PCR. Amplification products were analyzed on a 1.5% agarose gel. M: DNA Markers VI, Boehringer Mannheim. (Data kindly provided by D.J. MacKenzie, Centre for Plant Health, Agriculture and Agri-Food Canada and M.A. McLean, Euro Nursery & Vineyard (West) Inc., Sidney, Canada.)|Formaldehyde agarose gel and northern blot of total RNA (7 µg) from indicated tissues of tomato. Blot was hybridized with a 32P-labeled DNA probe for the tomato histone H4 gene. M: 0.24-9.5 kb RNA ladder. (Plant tissues and histone H4 cDNA clone kindly provided by K. Theres, MPI for Breeding Research, Cologne, Germany.)|
Performance
The RNeasy Plant Mini Kit is ideal for isolation of total RNA from a wide variety of plant and fungal samples with sample sizes of 10–100 mg tissue, or 100–1 x 107 cells (see table "Selected samples processed with the RNeasy Plant Mini Kit"). The binding capacity of the spin-columns is up to 100 µg RNA. The typical yield from 100 mg plant tissue is between 25 µg and 65 µg of RNA, although amounts of RNA can vary due to the developmental stage and growth conditions of the samples (see table "Yield from 100 mg tissue").
Anemone sp. Arabidopsis thaliana1 Begonia sp. Beta vulgaris (sugar beet)2 Chlamydomonas reinhardii (unicellular alga) Chrysanthemum Clarkia spp.3 Daucus carota (carrot)4 Diascia sp. Dendranthema sp. Euglena gracilis (unicellular alga) Funaria hygrometrica (moss) Hordeum vulgare (barley) Lycopersicon esculentum (tomato)5 Malus sp. (apple)6 Nicotiana tabacum (tobacco) Oryza sativa (rice) Pelargonium sp. (geranium) Petroselinum crispum (parsley)7 Pisum sativum (pea) Prunus sp. (cherry)6 Ranunculus sp. Ribes nigrum (black currant) Riccia fluitans (liverwort) Sinapis arvensis (mustard) Solanum tuberosum (potato)8 Spinacia oleracea (spinach) Surfinia sp. Triticum aestivum (wheat) Vitis sp. (grapevine)6 Zea mays (maize) |
Acremonium crysogenum9 Fusarium avenaceum9 |
| Arabidopsis leaves |
35 µg |
| Maize leaves |
25 µg |
| Tomato leaves |
65 µg |
| Tobacco leaves |
60 µg |
The purified plant RNA, eluted in 30–100 µl volume, includes viral RNA (see figure "Detection of viruses"). All contaminants, such as polysaccharides, are removed, and the eluted RNA is ready for all downstream applications (see figure "Tissue specificity of histone H4 expression").
Principle
The RNeasy Plant Mini Kit provides QIAshredder columns for homogenization and filtration of viscous plant or fungal lysates by microcentrifugation in combination with RNeasy Mini spin columns for RNA purification. RNeasy technology simplifies total RNA isolation by combining the stringency of guanidine-isothiocyanate lysis with the speed and purity of silica-membrane purification. RNeasy Kits provide the highest-quality RNA with minimum copurification of DNA. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using convenient on-column DNase treatment during the RNeasy procedure.
Procedure
Samples are first lysed and then homogenized in the QIAshredder columns. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the RNeasy silica membrane. RNA binds, and all contaminants are efficiently washed away. Pure, concentrated RNA is eluted in water (see flowchart " RNeasy Plant Mini procedure").
Applications
RNA purified with RNeasy technology is high-quality and is ideal for use in all applications. Downstream applications include:
- Northern, dot, and slot blotting
- End-point RT-PCR
- Quantitative, real-time RT-PCR
- Array analysis
- Poly A+ RNA selection
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RNeasy Mini Kit - 動物細胞、動物組織、バクテリア、酵母からのトータルRNA精製およびRNAクリーンアップ用
RNeasy Protect Mini Kit - 採取した動物組織からのRNAの迅速な安定化と続くトータルRNA精製
RNeasy Plant Mini Kit - 植物および糸状菌類からのトータルRNA精製用
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RNeasy Mini Kit - For purification of total RNA from animal cells, animal tissues, bacteria, and yeast, and for RNA cleanup; RNeasy Protect Mini Kit - For immediate stabilization of RNA in harvested animal tissues and subsequent total RNA purification; RNeasy Plant Mini Kit - For purification of total RNA from plants and filamentous fungi
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Download Safety Data Sheets for QIAGEN product components.
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画像
ウイルス検出
トータルRNAは、各種植物組織(感染したリンゴ(1)、サクランボ(2)、ナシ(3)およびブドウ(4~8)の芽、クロカラントの葉(9、10)、感染したジャガイモの休眠塊茎(11、12))から、RNeasyプロトコールの改良法を用いて精製した。ウイルスRNAはワンチューブRT-PCRにより増幅した。増幅産物は1.5%アガロースゲルで解析した。M:DNAマーカーVI、Boehringer Mannheim(データ提供:D.J.MacKenzie, Centre for Plant Health, Agriculture and Agri-Food Canada and M.A. McLean, Euro Nursery & Vineyard (West) Inc., Sidney, Canada)。
ヒストンH4発現の組織特異性
表記のトマト組織由来のトータルRNA(7 µg)のホルムアルデヒドアガロースゲルおよびノーザンブロット。ブロットは、トマトのヒストンH4遺伝子に対する32P標識DNAプローブによりハイブリダイズされた。M:0.24~9.5kb RNAラダー(植物組織およびヒストンH4 cDNAクローン提供:K. Theres, MPI for Breeding Research, Cologne, Germany)。
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