RNeasy Mini Kit

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").

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

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.

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.

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