For purification of up to 1 mg total RNA from cells, tissues, and yeast
Fast procedure delivering high-quality total RNA in minutes
Ready-to-use RNA for high performance in any downstream application
Consistent RNA yields from medium amounts of starting material
No phenol/chloroform extraction
No CsCl gradients, no LiCl or ethanol precipitation
The RNeasy Midi Kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane RNeasy spin columns with a binding capacity of 1 mg RNA. Tissue samples can be conveniently stabilized using RNAlater RNA Stabilization Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system. For smaller or larger samples, the RNeasy Micro Kit (spin column binding capacity of up to 45 µg RNA), the RNeasy Mini Kit (spin-column binding capacity of 100 µg RNA), and RNeasy Maxi Kit (spin-column binding capacity of 6 mg RNA) are also available.
The RNeasy Midi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
RNeasy Midi procedure.
Total RNA purified with the RNeasy Midi Kit is of high quality and is suitable for many downstream applications. Protocols are also included for cleanup of partially purified RNA, in vitro transcripts, and RNA from enzymatic reactions. An additional buffer is required (composition provided) for isolation of cytoplasmic RNA from eukaryotic cells. Lyticase, zymolase, or glass beads (required for yeast samples) are not provided. Amounts of RNA isolated from samples can vary due to the developmental stage, species, and growth conditions of the sample source. Since the RNeasy procedure enriches for RNA species >200 nt, RNA yield does not include 5S rRNA, tRNAs, or other low-molecular-weight RNAs.
RNeasy Midi spin column.
The RNeasy Midi spin column contained in the RNeasy Midi Kit.
Total RNA purified with the RNeasy Midi Kit is of high quality and is suitable for many downstream applications (see figure "High-quality RNA from a variety of samples"). Total RNA is easily purified with the RNeasy Midi Kit from animal or human cells, animal or human tissues, and yeast (seetable).
Total RNA yields obtained with the RNeasy Midi Kit
Average yield (µg)
Animal cells LMH HeLa COS-7 Lymphocytes (unstimulated)
7 x 107 7 x 107 3 x 107 1 x 108
850 1000 950 50
Mouse tissue Liver Lung Spleen
200 mg 100 mg 200 mg
700 130 600
Yeast cells S. cerevisiae
2 x 108
The RNeasy Midi Kit allows efficient purification of total RNA from medium amounts of starting material. 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.
With the RNeasy Midi Kit, total RNA is easily purified from 5 x 106 to 1 x 108 animal or human cells, 20–250 mg animal or human tissues, and 2 x 107 to 5 x 108 yeast cells. Samples are first lysed and then homogenized. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the RNeasy silica membrane (see figure "RNeasy Midi spin column"). RNA binds (up to 1 mg capacity), and all contaminants are efficiently washed away. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using a convenient on-column DNase treatment. Pure, concentrated RNA is eluted in 300–500 µl water (see figure "RNeasy Midi procedure"). A variety of special application protocols is also available.
RNA purified with RNeasy technology has A260/280 ratios of 1.9–2.1 (measured in 10 mM Tris·Cl, pH 7.5) and is ideal for use in all applications. Downstream applications include:
Northern, dot, and slot blotting
Quantitative, real-time RT-PCR
Poly A+ RNA selection
PCR, qPCR, real-time RT-PCR, microarray
Main sample type
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein