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.
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RNeasy Midi Kit (10)
10 RNeasy Midi Spin Columns, Collection Tubes (15 ml), RNase-free Reagents and Buffers
75142
$145.00
RNeasy Midi Kit (50)
50 RNeasy Midi Spin Columns, Collection Tubes (15 ml), RNase-free Reagents and Buffers
75144
$655.00
The RNeasy Midi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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RNeasy Midi procedure.|High-quality RNA from a variety of samples.|RNeasy Midi spin column.|
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.|Formaldehyde agarose gel and northern blot of total RNA purified with the RNeasy Maxi Kit. Total RNA (10 µg) isolated from each source was loaded per lane. All tissues were from mouse. Yeast: Saccharomyces cerevisiae; E. coli strain: HB101. 32P-labeled probes recognized (G) GAPDH; (E) translation elongation factor EF-1α; and (O) outer membrane protein A sequences. (E and O were kindly provided by P. Philippsen, University of Basel, Switzerland and U. Henning, Max Planck Institute of Biology, Tübingen, Germany, respectively.) B. subtilis was not probed. M: 0.24–9.5 kb RNA ladder. 7.5 kb band (indicated) in embryo, Huh7, and HeLa cell lanes is a nuclear precursor RNA.|The RNeasy Midi spin column contained in the RNeasy Midi Kit.|
Performance

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 (see table).

Total RNA yields obtained with the RNeasy Midi Kit
Source Starting materialAverage 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

450
 

Principle

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.

Procedure
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.
Applications

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
  • End-point RT-PCR
  • Quantitative, real-time RT-PCR
  • Array analysis
  • Poly A+ RNA selection
Feature
Specifications
Applications PCR, qPCR, real-time RT-PCR, microarray
Elution volume 300–500 µl
Format Spin column
Main sample type Tissue, cells
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 20–250 mg
Technology Silica technology
Time per run or per prep <1 hour
Yield Varies

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Kit Handbooks
1
RNeasy Midi Kit RNeasy Maxi Kit For total RNA isolation from animal cells animal tissues bacteria yeast whole blood and for RNA cleanup RNeasy Protect Midi Kit RNeasy Protect Maxi Kit For stabilization and protection of RNA in tissue For isolation of t
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