RNeasy Midi Kit
For purification of up to 1 mg total RNA from cells, tissues, and yeast
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.
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).
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.
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.
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:
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