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 RNAlater 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.
250 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), RNase-free Reagents and Buffers
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.
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.
RT-PCR of RNA from as few as 100 cells.
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.
RNeasy Mini spin column.
The RNeasy Mini spin column contained in the RNeasy Mini Kit.
Total RNA yields obtained with the RNeasy Mini Kit
Source
Starting material
Average yield (µg)
Animal cells LMH
HeLa
COS-7
Lymphocytes (unstimulated)
1 x 106 1 x 106 1 x 106 1 x 106
12
15
35
0.5
Mouse tissue Liver
Lung
Spleen
10 mg
10 mg
10 mg
40
10
35
Yeast cells S. cerevisiae
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 107 animal or human cells, 0.5–30 mg animal or human tissues, and <5 x 107 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:
