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RNeasy Lipid Tissue Mini Kit

For purification of up to 100 µg total RNA from fatty tissues and other types of tissue
  • Optimized lysis conditions for fatty tissues and other tissues
  • High yields of total RNA without phenol contamination
  • High-quality RNA for all downstream applications
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. 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.
Cat No./ID: 74804
RNeasy Lipid Tissue Mini Kit (50)
$466.00
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50 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), QIAzol Lysis Reagent, RNase-free Reagents and Buffers
The RNeasy Lipid Tissue Mini 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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High yields of RNA without phenol carryover.
RNA was isolated from 10 mg rat brain tissue using the RNeasy Lipid Tissue Mini Kit (RNeasy Lipid), a standard silica-gel-membrane procedure (Std. silica), or a phenol/guanidine-based reagent (PhOH/Gdn), following supplier's instruction. [A] Formaldehyde agarose gel analysis shows high yields of RNA using the RNeasy Lipid Tissue Mini Kit. Using the RNeasy and other silica-based methods, small RNAs (such as 5.8S rRNA, 5S rRNA, and tRNAs) are selectively excluded. [B] Absorbance spectrum shows contaminants and phenol when using the phenol/guanidine-based reagent.
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RNeasy Lipid Tissue Mini procedure.

The convenient RNeasy Lipid Tissue protocol integrates QIAzol phenol/guanidine-based lysis with silica-membrane 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. RNA partitions to the upper, aqueous phase, while DNA partitions to the interphase and proteins to the lower, organic phase or the interphase. Ethanol is added to the aqueous phase and the sample is applied to the RNeasy spin column.

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Real-time analysis of high-quality RNA.

Total RNA was isolated from 100 mg rat brain or adipose tissues using the RNeasy Lipid Tissue Mini Kit. Real-time, quantitative RT-PCR was carried out using QuantiTect Probe RT-PCR Kit with the indicated amounts of total RNA (in duplicate) and primers and probe specific for the c-jun or alpha-actin genes.

Performance
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").
Principle

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.

Procedure
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").
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.

Features
Specifications
Applications PCR, real-time PCR, microarray
Elution volume 30–100 µl
Format Spin column
Main sample type Fatty tissue samples
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 10–100 mg
Technology Silica technology
Time per run or per prep 45 minutes
Yield 2.4–10 mg

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Kit Handbooks (2)
For purification of total RNA from fatty tissues and all other types of tissue
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脂肪組織、脳、その他の脂肪性動物組織からのトータルRNA分離
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