Further information:
RNeasy Stabilization and Total RNA
Isolation System
For total RNA isolation from plants and
fungi
| Format: | Mini spin columns with 1.5 ml and 2 ml collection tubes |
| Sample source: | Plant and fungal cells or tissue |
| Sample size: | 10每100 mg tissue, 100 每 1 x 107 cells* |
| Preparation time: | 30 minutes |
| Binding capacity: | Up to 100 µg RNA |
| Yield (100 mg tissue)† | |
| Arabidopsis leaves: | 35 µg |
| Maize leaves: | 25 µg |
| Tomato leaves: | 65 µg |
| Tobacco leaves: | 60 µg |
| Elution volume: | 30每100 µl |
* Please call QIAGEN Technical Services
for guidelines to scale up the procedure.
† Amounts can vary due to developmental stage, growth conditions used,
etc. Since the RNeasy procedure enriches for RNA species >200 nt, RNA yield
does not include 5S rRNA, tRNAs, or other low-molecular-weight RNA species.
The RNeasy Plant Mini Kit is ideal for isolation of total RNA, including viral RNA, from a wide variety of plant and fungal samples (see figure "Detection of Viruses from a Variety of Plant Species by RT-PCR" and table "Selected samples processed with RNeasy Plant Mini Kit"). This kit provides QIAshredder columns for homogenization and filtration of viscous plant or fungal lysates by microcentrifugation in combination with RNeasy mini columns for RNA purification. 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").
Selected samples processed with RNeasy Plant Mini Kit
|
Plants |
|
| 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) Filamentous fungi Acremonium crysogenum9Fusarium avenaceum9 |
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) |
Young leaves or needles, except as indicated: 1 Flowers, buds, siliques, roots; 2 roots; 3 petals, stamen, stigma, sepals, style; 4 cultured cells; 5 mature flower, expanded leaves, shoot tip, stem, green fruit; 6 budwood; 7 seedlings; 8 dormant tuber; 9 mycelium
Detection of Viruses from a Variety of Plant Species by RT-PCR
Total RNA was isolated from budwood obtained from infected apple (1), cherry (2), pear (3), and grapevine (4每8), leaf tissue from black currant (9, 10), and dormant tuber tissue from infected potato (11, 12) using a modification of the RNeasy protocol (Ref. 1). 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.)
Tissue Specificity of Histone H4 Expression
 |
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 (Ref. 2). M: 0.24每9.5 kb RNA ladder. (Plant tissues and histone H4 cDNA clone kindly provided by K. Theres, MPI for Breeding Research, Cologne, Germany.) |
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 the RNase-Free DNase Set for convenient on-column DNase treatment during the RNeasy procedure.
RNeasy Mini Handbook 6/01 (PDF version, 351 KB)