RNeasy 96 Universal Tissue Kit

For 96-well purification of total RNA from any tissue

High yields of total RNA from all tissues
Streamlined QIAzol lysis and RNeasy 96 purification
Pure, high-quality RNA without phenol contamination
High-performance RNA for all downstream applications

The RNeasy 96 Universal Tissue Kit enables total RNA purification in 96-well format from any tissue. Tissues are efficiently lysed and homogenized in QIAzol Lysis Reagent, and high-quality RNA is purified using silica-membrane RNeasy 96 plates. Manual purification is carried out using the QIAGEN 96-Well-Plate Centrifugation System and, optionally, the QIAvac 96 vacuum manifold. Automated purification using the related RNeasy 96 Universal Tissue 8000 Kit is performed on the BioRobot Universal System. Tissue samples can be conveniently stabilized using Allprotect Tissue Reagent, and efficiently disrupted using the TissueLyser II. Nonfatty tissues can be stabilized using the RNAprotect Tissue Reagent.

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The RNeasy 96 Universal Tissue Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

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RNeasy 96 Universal Tissue procedure.|High quality total RNA from rat liver and muscle tissue.|High-quality RNA from skin tissue.|High-quality RNA from adipose tissue.|Real-time analysis of high-quality RNA from rat brain.|High-quality RNA from liver tissue.|High-quality RNA from intestine tissue.|

|Total RNA was purified from the indicated rat tissues using the RNeasy 96 Universal Tissue Kit and analyzed by formaldehyde agarose gel electrophoresis. The starting materials were 25 mg of RNAlater-stabilized liver and muscle tissue.|Total RNA was purified from 50 mg rat skin tissue using the RNeasy 96 Universal Tissue Kit and analyzed on the Agilent 2100 Bioanalyzer. |Total RNA was purified from 100 mg rat adipose tissue using the RNeasy 96 Universal Tissue Kit and analyzed on the Agilent 2100 Bioanalyzer.|Total RNA was purified from 50 mg samples of rat brain using the RNeasy 96 Universal Tissue Kit. Twenty-four purified samples, with an average of 90 ng RNA per reaction, were analyzed by real-time RT-PCR on the ABI PRISM 7900HT Sequence Detection System, using the QuantiTect Probe RT-PCR Kit with primers and probe specific for the c-jun gene.|Total RNA was purified from 25 mg rat liver tissue using the RNeasy 96 Universal Tissue Kit and analyzed on the Agilent 2100 Bioanalyzer.|Total RNA was purified from 50 mg rat intestine tissue using the RNeasy 96 Universal Tissue Kit and analyzed on the Agilent 2100 Bioanalyzer.|

Performance

Compared with standard silica-membrane procedures, the RNeasy 96 Universal Tissue Kit provides higher yields of total RNA for all tissue types (see table).

Typical total RNA yields using the RNeasy 96 Universal Tissue Kit
Tissue	RNA yield* (µg per 10 mg of tissue)
Kidney	5–40
Liver	15–80
Lung	5–15
Heart	5–25
Muscle	5–35
Brain	5–20
Adipose tissue	0.5–2.5
Spleen	15–100
Intestine	10–60
Skin	2–5

High-quality RNA can be isolated from tissues preserved either by freezing or by stabilization with RNAprotect at room temperature (see figure "High-quality total RNA from rat liver and muscle tissue"). Intact RNA purified from difficult-to-lyse fibrous and fatty tissues is suitable for downstream applications (see figures "High-quality RNA from skin / adipose / liver / intestine tissue" and "Real-time analysis of high-quality RNA from rat brain").

Principle

The RNeasy 96 Universal Tissue procedure enables high-throughput purification of RNA from any animal or human tissue sample. Compared with standard silica-membrane procedures, the RNeasy 96 Universal Tissue Kit provides higher yields of total RNA for all tissue types. The RNeasy procedure enriches for RNA >200 bases long, providing a yield of total RNA that does not include 5.8S rRNA, tRNA, and other low-molecular weight RNAs, which make up 15–20% of total cellular RNA.

Procedure

The optimized RNeasy 96 Universal Tissue procedure integrates TissueLyser/QIAzol processing with RNeasy 96 purification in an easy-to-follow protocol (see flowchart "RNeasy 96 Universal Tissue procedure"). Efficient high-throughput disruption using the TissueLyser II provides parallel disruption and homogenization, with lysis of up to 192 samples in QIAzol Lysis Reagent. Protocol steps are performed manually using the QIAGEN 96-Well-Plate Centrifugation System, with optional use of the QIAvac 96 vacuum manifold. For automated processing, see the RNeasy 96 Universal Tissue 8000 Kit.

Applications

The combination of QIAzol and RNeasy technologies results in highly pure RNA without phenol carryover. RNA purified using the RNeasy 96 Universal Tissue Kit is suitable for all downstream applications, including real-time RT-PCR and array analysis.

Feature	Specifications
Applications	PCR, qPCR, real-time PCR
Elution volume	45–70 µl
Format	96-well plate
Main sample type	Tissue samples
Processing	Manual (centrifugation or vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	RNA
Sample amount	25–100 mg
Technology	Silica technology
Yield	Varies

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