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. The kit can also be used in combination with a
TissueRuptor or TissueLyser system, which provide efficient disruption and homogenization of plant samples.
50 RNeasy Mini Spin Columns, 50 QIAshredder Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
The RNeasy Plant Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
RNeasy Plant Mini procedure.
Detection of viruses.
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.)
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. 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").
Selected samples processed with the RNeasy Plant Mini Kit
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
Features
Specifications
Applications
PCR, qPCR, real-time RT-PCR, microarray
Elution volume
30–100 µl
Format
Spin column
Main sample type
Plant samples
Processing
Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
