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RNeasy Mini, Midi, and Maxi Kits

For total RNA isolation from a wide variety of sources RNeasy Protect
Mini Kit
RNeasy Protect Midi Kit RNeasy Protect Maxi Kit
Format: Mini columns with 1.5 ml and 2 ml collection tubes Midi columns with 15 ml collection tubes Maxi columns with 50 ml collection tubes Sample size
0.530 mg 20250 mg 150 mg 1 g Volume of
Stabilization Reagent:
50 ml (50-prep kit)
250 ml (250-prep kit) 20 ml (10-prep kit) or
100 ml (50-prep kit) 100 ml (12-prep kit) Binding capacity: Up to 100 g RNA Up to 1 mg RNA Up to 6 mg RNA Yield: Varies depending on sample source. See table Elution volume: 30100 l 300500 l 8002400 l RNeasy Mini, Midi, and Maxi Kits allow efficient purification of total RNA from very small (see figure "RT-PCR of RNA from =100 Cells") to very large amounts of starting material. Total RNA is easily purified from animal cells or tissues, Gram-positive or Gram-negative bacteria,* or yeast (See table "Total RNA Yields Obtained with RNeasy Mini, Midi, and Maxi Kits" and figure "High-Quality RNA from a Variety of Samples"). Protocols are also included for isolation of cytoplasmic RNA from eukaryotic cells as well as for cleanup of partially purified RNA, in vitro transcripts, and RNA from enzymatic reactions. A variety of special application protocols, including a protocol for vacuum processing of RNeasy mini columns, are also available. QIAshredder Homogenizers for convenient cell-lysate homogenization are available as an accessory.

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.

* Lysozyme (required for bacterial samples) is not provided.
Lyticase, zymolase, or glass beads (required for yeast samples) are not provided.
Additional buffer required. The buffer composition is provided in the protocols.

High-Quality RNA from a Variety of Samples Formaldehyde agarose ge l and northern blot of total RNA purified with the RNeasy Maxi Kit. 10 g of total RNA isolated from each source was loaded per lane. All tissues were from mouse. Yeast: Saccharomyces cerevisiae; E. coli: HB101. 32P-labeled probes recognized (G) GAPDH; (E) translation elongation factor EF-1a ; 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, Tbingen, Germany, respectively.) B. subtilis was not probed. M: 0.249.5 kb RNA ladder. 7.5 kb band (indicated) in embryo, Huh7, and HeLa cell lanes is a nuclear precursor RNA.
RT-PCR of RNA from >100 Cells RT-PCR of total RNA isolated with the RNeasy Mini Kit from the indicated numbers of HeLa cells. 10 l (1/5) of eluate was digested with RNase-free DNase and reverse transcribed with oligo-dT primer. 2.5 l (1/20) of the cDNA mix was used in 50 l PCR. A 452 bp fragment of GAPDH was amplified. C: negative control; C+: positive control; M: 100 bp ladder.
Total RNA Yields Obtained with RNeasy Mini, Midi, and Maxi Kits Amounts can vary due to developmental stage, species, 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 RNAs.
* Using the specialized protocol for heart, muscle, and skin tissue, included in handbook. Proteinase K (required for this protocol) is not provided.
Higher yields can be obtained from stabilized bacteria using RNeasy Protect Bacteria Kits.< br> * Requires use of a centrifuge capable of attaining 30005000 x g equipped with a swing-out rotor for 15 ml (Midi) or 50 ml (Maxi) centrifuge tubes.



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