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RNeasy Maxi Kit

For purification of up to 6 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 large amounts of starting material
  • No phenol/chloroform extraction
  • No CsCl gradients, no LiCl or ethanol precipitation
The RNeasy Maxi Kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane RNeasy spin columns with a binding capacity of 6 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 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 Midi Kit (spin-column binding capacity of 1 mg RNA) are also available.
Cat No./ID: 75162
RNeasy Maxi Kit (12)
12 RNeasy Maxi Spin Columns, Collection Tubes (50 ml), RNase-free Reagents and Buffers
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RNeasy Maxi procedure.

RNeasy Kits provide the highest-quality RNA with minimum copurification of DNA. An additional buffer (composition provided) is required 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.

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High-quality RNA from a variety of samples.
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; Strain of E. coli: 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.
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RNeasy Maxi spin column.
The RNeasy Maxi spin column contained in the RNeasy Maxi Kit.
Performance

Total RNA purified with the RNeasy Maxi 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 Maxi Kit from large amounts of starting material including animal or human cells, animal or human tissues, and yeast cells (see table).

Total RNA yields obtained with the RNeasy Maxi Kit
Source Starting materialAverage yield (mg)

Animal cells
LMH
HeLa
COS-7
Lymphocytes (unstimulated)


5 x 108
4 x 108
1.8 x 108
5 x 108


5.7
6.0
5.8
0.3

Mouse tissue
Liver
Lung
Spleen


1 g
0.5 g
1 g

3.6
0.6
3.2

Yeast cells
S. cerevisiae


1 x 109

2.4
 

Principle

The RNeasy Maxi Kit delivers efficient purification of total RNA from large 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. Total RNA is easily purified from animal or human cells, animal or human tissues, and yeast. Protocols are included in the kit for cleanup of partially purified RNA, in vitro transcripts, and RNA from enzymatic reactions. RNeasy purified nucleic acids can be used in any molecular assay or other downstream application.

Procedure
RNeasy technology simplifies total RNA isolation. Samples (up to 5 x 108 animal or human cells, 2.5 x 109 yeast cells, or 150 mg to 1 g tissue) 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 Maxi spin column"). RNA (up to 6 mg) binds to the membrane, and all contaminants are efficiently washed away. Residual amounts of DNA can be removed using the convenient on-column DNase treatment. Pure, concentrated RNA is eluted in 800–2400 µl water (see figure "RNeasy Maxi 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
Features
Specifications
Applications PCR, qPCR, real-time RT-PCR, microarray
Elution volume 800–2400 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 150 mg – 1 g
Technology Silica technology
Time per run or per prep 1 hour
Yield Varies

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Kit Handbooks (1)

For total RNA isolation from animal cells, animal tissues, bacteria, yeast, whole blood, and for RNA cleanup


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