RNeasy Lipid Tissue Mini Kit
For purification of up to 100 µg total RNA from fatty tissues and other types of tissue
The RNeasy Lipid Tissue Mini Kit includes QIAzol Lysis Reagent for lysing fatty tissues and other types of tissue, and RNeasy spin columns for purifying up to 100 µg of high-quality RNA. The kit can be automated using the QIAcube Connect. 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 larger samples, the RNeasy Lipid Tissue Midi Kit (spin-column binding capacity of 1 mg RNA) is also available.
The RNeasy Lipid Tissue protocol integrates QIAzol lysis with RNeasy RNA isolation for high yields of total RNA from fatty tissues with an average yield of RNA from 10 mg of brain of 10 µg, and 2.4 µg of RNA from a 10 mg sample of adipose tissue. RNeasy Lipid Tissue kits provide efficient isolation of high-quality total RNA without phenol carryover (see figure "High yields of RNA without phenol carryover"). The RNA from fatty tissues is suitable for downstream applications including real-time RT-PCR (see figure "Real-time analysis of high-quality RNA").
RNeasy Lipid Tissue Kits are optimized for use with fatty tissues, such as brain and adipose tissue. The convenient RNeasy Lipid Tissue protocol integrates optimized phenol/guanidine-based lysis with proven RNeasy purification for isolation of high yields of high-quality total RNA. The combination of organic extraction and chaotropic disruption contributes to the high lysis efficiency of QIAzol Lysis Reagent. 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 10–100 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 100 µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30–100 µl of RNase-free water water (see figure "RNeasy Lipid Tissue Mini 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.
fragment fix placeholder