RNeasy Lipid Tissue Midi Kit
For purification of up to 1 mg total RNA from fatty tissues and other types of tissue
The RNeasy Lipid Tissue Midi Kit includes QIAzol Lysis Reagent for lysing fatty tissues and other types of tissue, and RNeasy spin columns for purifying up to 1 mg of high-quality RNA. Tissue samples can be conveniently stabilized using RNAlater RNA Stabilization Reagent (nonfatty tissues only) or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system. For smaller samples, the RNeasy Lipid Tissue Mini Kit (spin-column binding capacity of 100 µg RNA) is also available.
The RNeasy Lipid Tissue Midi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
RNeasy Lipid Tissue Kits are optimized for use with fatty tissues with RNA yields from 100 mg of representative fatty tissues of 110 µg from brain, and 21 µg from adipose tissue. The RNeasy Lipid Tissue procedure ensures easy and efficient isolation of high-quality total RNA without phenol carryover (see figures "High-quality RNA" and "High yields of RNA without phenol carryover"). RNA purified with the kits from lipid tissue is suitable for real-time RT-PCR (see figure "Real-time analysis").
RNeasy Lipid Tissue Kits are optimized for use with fatty tissues, such as brain and adipose tissue. These kits integrate optimized phenol/guanidine-based lysis with proven RNeasy silica-membrane purification for isolation of high-quality total RNA. The combination of organic extraction and chaotropic disruption contributes to the high lysis efficiency of QIAzol Lysis Reagent, enabling use of larger amounts of fatty tissues in the RNeasy procedure for high yields of total RNA. Since the RNeasy procedures enrich for mRNA and other RNA species >200 nucleotides, the total RNA yield does not include 5S rRNA, tRNA, and other low-molecular-weight RNAs, which make up 15-20% of total cellular RNA.
Tissue samples 75–500 mg are homogenized in QIAzol Lysis Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the RNeasy spin column, where the total RNA up to 1 mg binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 150–500 µl RNase-free water water (see figure "RNeasy Lipid Tissue Midi procedure").
RNeasy Lipid Tissue Kits provide easy and efficient isolation of high-quality RNA for all downstream applications, including array analysis and real-time RT-PCR.
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