For purification of up to 100 µg 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 small amounts of starting material
- No phenol/chloroform extraction, no CsCl gradients, no LiCl or ethanol precipitation
The RNeasy Mini Kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane RNeasy spin columns with a binding capacity of 100 μg RNA. Tissue samples can be conveniently stabilized using RNA later RNA Stabilization Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system. RNA purification using the RNeasy Mini Kit can be fully automated on the QIAcube. For smaller and larger samples, the RNeasy Micro Kit (spin-column binding capacity of up to 45 µg RNA), the RNeasy Midi Kit (spin-column binding capacity of 1 mg RNA), and RNeasy Maxi Kit (spin-column binding capacity of 6 mg RNA) are also available.
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RNeasy Mini Kit (50)
50 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
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74104
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RNeasy Mini Kit (250)
250 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
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74106
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The RNeasy Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
RNeasy Mini procedure.|High-quality RNA from a variety of samples.|RT-PCR of RNA from as few as 100 cells.|RNeasy Mini spin column.|
Total RNA purified with the RNeasy Mini 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. 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.|RT-PCR of total RNA isolated with the RNeasy Mini Kit from the indicated numbers of HeLa cells. 10 µl (1/5) of eluate was digested with RNase-free DNase and reverse transcribed with oligo-dT primer. 2.5 µl (1/20) of the cDNA mix was used in 50 µl PCR. A 452 bp fragment of GAPDH was amplified. C-: negative control; C+: positive control; M: 100 bp ladder.|The RNeasy Mini spin column contained in the RNeasy Mini Kit.|
Performance
Total RNA is easily purified with the RNeasy Mini Kit from up to 30 mg animal or human tissues, from 100 to 1 x 107 animal or human cells, or yeast (see table; and figures "RT-PCR of RNA from as few as 100 cells" and "High-quality RNA from a variety of samples").
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Animal cells
LMH
HeLa
COS-7
Lymphocytes (unstimulated)
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1 x 106
1 x 106
1 x 106
1 x 106 |
12
15
35
0.5
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Mouse tissue
Liver
Lung
Spleen
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10 mg
10 mg
10 mg |
40
10
35 |
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Yeast cells
S. cerevisiae
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1 x 107 |
25 |
Principle
The RNeasy Mini Kit allows efficient purification of total RNA from small 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 Mini Kit, total RNA is easily purified from 10 to 1 x 10 7 animal or human cells, 0.5–30 mg animal or human tissues, and <5 x 10 7 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 Mini spin column"). RNA binds (up to 100 µg 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 30–100 µl water (see figure " RNeasy Mini 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
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Feature
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Specifications
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Applications
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Northern, dot, and slot blotting, end-point RT-PCR, quantitative, real-time RT-PCR, array analysis, next-generation sequencing
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Elution volume
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30–100 µl
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Format
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Spin column
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Main sample type
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Tissue, cells
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Processing
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Manual (centrifugation)
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Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
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Total RNA
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Sample amount
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0.5–30 mg
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Technology
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Silica technology
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Time per run or per prep
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20 minutes
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Yield
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Varies
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RNeasy Mini Kit - For purification of total RNA from animal cells, animal tissues, bacteria, and yeast, and for RNA cleanup; RNeasy Protect Mini Kit - For immediate stabilization of RNA in harvested animal tissues and subsequent total RNA purification; RNeasy Plant Mini Kit - For purification of total RNA from plants and filamentous fungi
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Up to 1 x 109 bacteria are disrupted and homogenized by bead-milling in a guanidine-thiocyanate-containing lysis buffer. After addition of ethanol, the sample is loaded onto an RNeasy Mini spin column. Total RNA binds to the RNeasy silica-membrane, contaminants are efficiently washed away, and high-quality RNA is eluted in RNase-free water.
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One milliliter of milk contains approximately 50,000-200,000 leukocytes (with an average of approximately 100,000 leukocytes). In a BVDV-infected animal up to 1-30% of the leukocytes may be infected with the virus. Each infected leukocyte typically contains 10-10,000 BVDV entities.
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