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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

S_1270_GEF_RNALT0191

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RNeasy Lipid Tissue Mini Kit (50)

Cat no. / ID.   74804

50 RNeasy Mini Spin Columns, Collection Tubes (1.5 ml and 2 ml), QIAzol Lysis Reagent, RNase-free Reagents and Buffers
CA$727.00
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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.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Optimized lysis conditions for fatty tissues and other tissues
  • High yields of total RNA without phenol contamination
  • High-quality RNA for all downstream applications

Product Details

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 RNAprotect Tissue Reagent (nonfatty tissues only) or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor or TissueLyser system.

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 Kit 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 Kit is 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 Kit provides easy and efficient isolation of high-quality RNA for all downstream applications, including NGS and real-time RT-PCR.

Supporting data and figures

Specifications

FeaturesSpecifications
Sample amount10–100 mg
Yield2.4–10 mg
TechnologySilica technology
ProcessingManual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinRNA
Elution volume30–100 µl
Main sample typeFatty tissue samples
Time per run or per prep45 minutes
FormatSpin column

Publications

Lipopolysaccharide activates an innate immune system response in human adipose tissue in obesity and type 2 diabetes.
Creely SJ; McTernan PG; Kusminski CM; Fisher fM; Da Silva NF; Khanolkar M; Evans M; Harte AL; Kumar S;
Am J Physiol Endocrinol Metab; 2006; 292 (3):E740-7 2006 Nov 7 PMID:17090751
Chronic ethanol feeding to rats decreases adiponectin secretion by subcutaneous adipocytes.
Chen X; Sebastian BM; Nagy LE;
Am J Physiol Endocrinol Metab; 2006; 292 (2):E621-8 2006 Oct 17 PMID:17047161
Targeting an E2F site in the mouse genome prevents promoter silencing in quiescent and post-mitotic cells.
Tavner F; Frampton J; Watson RJ;
Oncogene; 2006; 26 (19):2727-35 2006 Oct 30 PMID:17072340
Paternally biased X inactivation in mouse neonatal brain.
Wang X; Soloway PD; Clark AG;
Genome Biol; 2010; 11 (7):R79 2010 Jul 27 PMID:20663224
A study of alternative splicing in the pig.
Nygard AB; Cirera S; Gilchrist MJ; Gorodkin J; Jørgensen CB; Fredholm M;
BMC Res Notes; 2010; 3:123 2010 May 5 PMID:20444244

FAQ

How should RNeasy Kits be stored and how long are they stable?
RNeasy Mini, Midi and Maxi Kits should be stored dry at room temperature (15 to 25°C). The RNeasy MinElute Spin Columns of the RNeasy Micro Kit and RNeasy MinElute Cleanup Kit should be stored at 4°C. RNeasy Kits are stable for at least 9 months under these conditions.
FAQ-103
How do you ensure that RNeasy buffers are RNase-free?

Our RNeasy buffers are subjected to stringent quality-control tests to ensure that they are indeed RNase-free. Buffer RPE concentrate and RNase-free water are tested for absence of RNases by incubating 4 µg of total HeLa-RNA in these solutions for 3 hours at 37°C, followed by monitoring RNA integrity via denaturing agarose gel electrophoresis and ethidium bromide staining.

Buffer RLT and Buffer RW1 are inherently RNase-free, since the buffers themselves inactivate RNases during the RNeasy procedure.

FAQ-113
What is the difference between disruption and homogenization in the RNeasy System?

Disruption:

Complete disruption of cell walls and plasma membranes of cells and organelles is absolutely required to release all RNA contained in a sample. Different samples require different methods to achieve complete disruption. Please refer to the section 'Disruption and homogenization of starting materials' in the RNeasy Mini Handbook. Incomplete disruption results in significantly reduced RNA yields.

Homogenization:

Homogenization is necessary to reduce the viscosity of the cell lysates produced by disruption. Homogenization shears the high-molecular-weight genomic DNA and other high-molecular-weight cellular components to create a homogeneous lysate. Incomplete homogenization results in genomic DNA contamination, and inefficient binding of RNA to the RNeasy membrane resulting in reduced yields.

FAQ-139
Are RNeasy spin columns sold separately?
At this time, RNeasy spin columns are not sold separately.
FAQ-159
What is the maximum binding capacity of RNeasy spin columns?
The maximum binding capacity of the RNeasy Mini spin columns is 100 ug RNA. It is 1 mg for RNeasy Midi columns and 6 mg for RNeasy Maxi columns. The maximum RNA binding capacity of the RNeasy MinElute spin columns is 45 µg.
FAQ-290