Principle

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 purification for isolation of high-quality total RNA without phenol carryover (see figure "High yields of RNA without phenol carryover").

Specifications of the RNeasy Lipid Tissue Mini Kit

Specification	Details
Format	Mini spin columns with 1.5 ml and 2 ml collection tubes
Sample size	10–100 mg
Preparation time	45 minutes
Binding capacity	Up to 100 µg RNA
Average yield* from 10 mg tissue	Brain: 10 µg Adipose tissue: 2.4 µg
Elution volume	30–100 µl

* Amounts can vary due to species, developmental stage, etc. 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.

The convenient RNeasy Lipid Tissue protocol integrates QIAzol lysis with RNeasy RNA isolation for high yields of 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.

RNeasy Lipid Tissue technology simplifies total RNA isolation by combining the stringency of phenol/guanidine-based lysis with the speed and purity of silica-membrane purification. There is no need for centrifugation through CsCl cushions or precipitation with LiCl or alcohol. RNeasy sample preparation technology is fully licensed, allowing RNeasy purified nucleic acids to be used in any molecular assay or other downstream application without risk of patent infringement.

Procedure

Tissue samples are homogenized in QIAzol Lysis Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. RNA partitions to the upper, aqueous phase, while DNA partitions to the interphase and proteins to the lower, organic phase or the interphase.

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 binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in RNase-free water water.

Applications

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 (see figure "Real-time analysis of high-quality RNA from fatty tissues").

A range of optimized kits for RNA purification from tissue samples are available. Use the ProductFinder to choose the right kit.