RNeasy Midi Kit

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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Product		Cat. no.	List price:
RNeasy Midi Kit (10) 10 RNeasy Midi Spin Columns, Collection Tubes (15 ml), RNase-free Reagents and Buffers		75142	Inquire
RNeasy Midi Kit (50) 50 RNeasy Midi Spin Columns, Collection Tubes (15 ml), RNase-free Reagents and Buffers		75144	Inquire

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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 material	Average yield (µg)
Animal cells LMH HeLa COS-7 Lymphocytes (unstimulated)	7 x 10⁷7 x 10⁷3 x 10⁷1 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 A_260/280 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

Brochures & Guides

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	(EN) - Analyzing Gene Expression and Regulation	Show details
	Product Profile - RNeasy® Mini, Midi and Maxi Kits	Show details
	(EN) - Quality RNA	Show details

FAQs

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	How do I safely inactivate biohazardous flow-through material? FAQ ID -12	View
	How can I avoid little or no RNA yields when using an RNeasy Kit? FAQ ID -28	View
	How should I quantify RNA isolated with RNeasy Kits? FAQ ID -32	View
	Why does my isolated RNA have a low OD 260/280 ratio? FAQ ID -97	View
	Why do I have to add beta-mercaptoethanol (beta-ME) to lysis Buffer RLT of the RNeasy Kits? FAQ ID -101	View
	How should RNeasy Kits be stored and how long are they stable? FAQ ID -103	View
	How do you ensure that RNeasy buffers are RNase-free? FAQ ID -113	View
	How can I isolate RNA from 1 gram of plant tissue? FAQ ID -128	View
	What is the difference between disruption and homogenization in the RNeasy System? FAQ ID -139	View
	How can I obtain DNA-free RNA using an RNeasy Midi or Maxi Kit? FAQ ID -143	View
	Are RNeasy spin columns sold separately? FAQ ID -159	View
	How can I check for purity of RNA isolated using RNeasy Kits? FAQ ID -1023	View
	How can I check the integrity of RNA purified using RNeasy Kits? FAQ ID -1024	View
	Which kit should I use for RNA isolation from Cartilage? FAQ ID -1026	View
	Do I have to discard Buffer RLT with beta-Mercaptoethanol (ß-ME) added to it after 1 month of storage? FAQ ID -1037	View
	What is the difference between Buffers RLT and RLT Plus? FAQ ID -1043	View
	How can I ensure complete genomic DNA removal when using the RNase-Free DNase Set? FAQ ID -1087	View
	Can acetone be used for precipitation of protein from Buffer RLT lysates generated with RNeasy Kits? FAQ ID -1164	View
	What is the maximum volume of RNA in solution that can be used with the QuantiTect Whole Transcriptome Kit? FAQ ID -1616	View
	Have you observed co-amplification of genomic DNA from RNA templates used in the QuantiTect Whole Transcriptome Procedure? FAQ ID -1619	View
	Can I clean up my DNase treated RNA samples using RNeasy columns? FAQ ID -286	View
	What is the maximum binding capacity of RNeasy spin columns? FAQ ID -290	View
	What is the composition of Buffer RLT? FAQ ID -2793	View
	What is the composition of Buffer RW1? FAQ ID -2796	View
	What is the composition of Buffer RPE? FAQ ID -2797	View
	What is the composition of Buffer RDD? FAQ ID -2800	View
	What kits does QIAGEN offer to extract RNA from whole blood? FAQ ID -304	View
	Can I buy the RNeasy Mini columns, RNeasy MinElute columns, RNeasy Midi columns or the RNeasy Maxi columns separately? FAQ ID - 3388	View
	Which kit should be used to extract RNA from adipose tissue, brain, and other fatty animal tissues? FAQ ID -467	View
	What has to be done to a RNA sample before loading it onto an Agilent Bioanalyzer? FAQ ID -528	View
	Is it possible to isolate both RNA and recombinant 6xHis-tagged protein from the same sample? FAQ ID -532	View
	Do you have a protocol for isolating RNA from yeast using RNeasy? FAQ ID -535	View
	Do you offer QIAshredder spin columns for the RNeasy Midi/Maxi Kit format? If not, what is the alternative? FAQ ID -560	View
	What do you recommend for isolating RNA from more than 100 mg of plant tissue? FAQ ID -570	View
	Can the QIAshredder Maxi Spin Columns from the DNeasy Plant Maxi Kit be used for RNeasy samples? FAQ ID -616	View
	Can the RNase-Free DNase Set be used for DNase digestions of RNA in solution? FAQ ID -619	View
	How can I improve my RNA yield from liver sample when using RNeasy Kits? FAQ ID -623	View
	How can I minimize degradation of RNA from my pancreas sample? FAQ ID -624	View
	Do you have a kit for RNA isolation from any kind of sample type? FAQ ID -627	View
	What is a QIAshredder? Is it sufficient for complete disruption and homogenization of my tissue sample? FAQ ID -631	View
	What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA? FAQ ID -728	View
	I ran my RNA out on an agarose gel and can see lots of bands similar to a ladder. Why? FAQ ID -745	View
	Is RNAprotect Bacteria Reagent compatible with the RNeasy Mini Kit? FAQ ID -797	View
	Do I need to use RNase inhibitors with the RNeasy Kits? FAQ ID -813	View
	What do you suggest to prevent degradation of RNA isolated from tissue with high amounts of RNases using RNeasy? FAQ ID -942	View
	Effects of low A260/A230 ratios in RNA preparations on downstream applications FAQ ID -2248	View
	I accidentally stored Buffer RDD of the RNase-Free DNase Set at -20°C. Will it still function?	View

Supplementary Protocols

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	Acetone precipitation of protein from Buffer RLT or Buffer RLT Plus lysates - (EN)	Show details

User-Developed Protocols

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	Purification of total RNA, including small RNAs, using the RNeasy® Midi Kit - (EN)	Show details

Kit Handbooks

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	RNeasy Midi/Maxi Handbook - (EN) 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	Show details

Quick-Start Protocols

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	RNeasy Midi Kit (EN)	Show details

Scientific Posters

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	Explore the RNA Universe! Poster for download	Show details

Safety Data Sheets

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	MSDS RNeasy Midi Kit (10)
	MSDS RNeasy Midi Kit (50)
	CofA

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