QIAwave RNA Mini Kit

For a more eco-friendly alternative to our standard kit for extracting total RNA from cells, tissues, and yeast.

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QIAwave RNA Mini Kit (250)

Cat. No. / ID: 74536

250 RNeasy Mini Spin Columns, Waste Tubes (2 ml), RNase-free Reagents and Buffers

KitKit component

QIAwave RNA Mini Kit

Waste Tubes

QIAwave RNA Mini Kit (250)は分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

RNA quality and performance identical to the RNeasy Mini Kit
61% less plastic and 38% less cardboard compared to the RNeasy Mini Kit
Reusable Waste Tubes made from 100% post-consumer recycled plastic
Buffer concentrates that use 86% less plastic than our standard buffer bottles

Product Details

The QIAwave RNA Mini Kit is an eco-friendlier version of our standard RNeasy Mini Kit. The QIAwave Kit uses 61% less plastic and 38% less cardboard than our standard kit and offers waste tubes made from 100% post-consumer recycled plastic that you can reuse throughout the procedure. QIAwave buffers also come as concentrates, reducing the amount of plastic by 86% per bottle. To save paper, there are no printed protocols in the kit. Instead, you can download the protocols you need from the resources list or by simply scanning the QR code inside the box lid. So, while the kit packaging and components of our QIAwave Kit may look different, it’s as easy to use as the RNeasy Mini Kit, and the chemistry and performance are identical.

Please be aware that you will need sterile glass bottles to store the reconstituted buffers.

In partnership with My Green Lab, we've also assessed the environmental impact of this kit. My Green Lab ACT (accountability, consistency, and transparency) environmental impact factor labels are designed to evaluate and score products on several sustainability criteria. These include:

manufacturing
responsible chemical management
sustainable content within products and packaging materials
disposal of the packaging at the end of life

Products are scored from 1 to 10 except for energy and water consumption, which are scored as 1 point per kWh or gallon, respectively. A low score means a lower environmental impact (see figures "QIAwave RNA Mini Kit ACT environmental impact factor label US, EU and UK").

The QIAwave RNA Mini Kit offers you fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane RNeasy Mini Spin Columns. Before extraction, you can stabilize tissue samples using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and to efficiently homogenize your tissue samples, you can use the TissueRuptor II, TissueLyser II or LT system.
If you have too many samples to prep manually, you can automate your sample extraction on the QIAcube Connect.

See figures

QIAwave RNA Mini Kit ACT environmental impact factor label US

QIAwave RNA Mini Kit ACT environmental impact factor label EU.

QIAwave RNA Mini Kit ACT environmental impact factor label UK.

Performance

The QIAwave RNA Mini Kit gives you highly reproducible yields of total RNA from animal or human cells, animal or human tissues, and yeast (see table “Total RNA yields obtained with RNeasy Mini Spin Columns” and figure " RT-PCR of RNA from as few as 100 cells").
The performance between our QIAwave RNA Mini Kit and RNeasy Mini Kit is identical because the chemistry is the same. We’ve also shown that both kits outperform competitors’ kits (see figure "QIAwave RNA Mini Kit performance").

Total RNA yields obtained with RNeasy Spin Mini Columns

Animal Cells
Source	Average Yield
	Mini	Mini	C
LMH	1 x 10⁵	12 µg	1.3 µg*
HeLa	1 x 10⁶	15 µg	1.6 µg*
COS-7	1 x 10⁶	35 µg	3.1 µg*
Lymphocytes (unstimulated)	1 x 10⁶	0.5 µg	-
Huh7	-	-	2.0 µg*
Jurkat	-	-	1.4 µg*
Mouse tissue
Liver	10 mg	40 µg	-
Lung	10 mg	10 µg	-
Spleen	10 mg	35 µg	-
Yeast cells
S. cerevisiae	1 x 10⁷	25 µg	-

*Amounts can vary due to developmental stage, growth conditions used, etc.

See figures

RT-PCR of RNA from as few as 100 cells.

QIAwave Kit performance.

Principle

The QIAwave RNA Mini Kit allows you to efficiently purify total RNA from samples such as:

microdissected tissues and fine needle aspirates
milligram amounts of fibrous tissues, including heart and muscle tissue
small numbers of cells down to a single cell (e.g., FACS sorted cells)

For microdissected FFPE tissues, we recommend using the RNeasy FFPE Kit. The QIAwave Kit simplifies total RNA isolation by combining the stringency of guanidine-isothiocyanate lysis with the speed and purity of silica-membrane purification. The QIAwave RNA Mini Kit also gives you high-quality RNA with minimal copurification of DNA.

Procedure

With the QIAwave Kit, you can purify RNA in four simple steps (see flowchart " QIAwave RNA Mini Procedure").

Lyse and homogenize samples (see table "Amount of starting material for the QIAwave RNA Mini Kit").
Add ethanol to the lysate to provide ideal binding conditions.
Load the lysate onto the RNeasy silica membrane. At this point, the RNA binds to the silica membrane and all contaminants are efficiently washed away. If your downstream applications are sensitive to very small amounts of DNA, you can remove any residual amounts of DNA using a convenient on-column DNase treatment.
Pure, concentrated RNA is then eluted in water.

In addition to our standard protocol, we also provide a variety of special application protocols.
If you have too many samples to process manually, you can automate the purification process on the QIAcube Connect. The QIAcube Connect allows you fully automate the entire purification procedure for up to 12 samples per run.
QIAwave buffers come as concentrates that you can easily reconstitute by adding water and/or ethanol; please check the handbook for details. QIAwave RNeasy Mini Spin Columns and Waste Tubes come in individual bags and need to be preassembled before you start the protocol. This takes a little extra time, but it does reduce plastic waste.

Amount of starting material for the QIAwave RNA Mini Kit

Amount of starting material	Mini
Cells	10 to 1 x 10⁷ animal or human cells
Tissue	0.5–30 mg animal or human tissues
Yeast	<5 x 10⁷ yeast cells

See figures

QIAwave RNA Mini procedure

Applications

RNA purified with QIAwave RNA technology has A260/280 ratios of 1.9–2.1 (measured in 10 mM Tris·Cl, pH 7.5) and is ideal for use in all applications. Downstream applications include:

RNA-seq
Quantitative, real-time RT-PCR
Real-time RT-PCR starting with as little as one cell
End-point RT-PCR
Northern, dot, and slot blotting
Array analysis
Poly A+ RNA selection

Supporting data and figures

RT-PCR of RNA from as few as 100 cells.

RT-PCR of total RNA isolated with RNeasy Mini Spin Columns from the indicated numbers of HeLa cells. 10 µl (1/5) of eluate was digested with RNase-free DNase and reverse transcribed with oligo-dT primer. 2.5 µl (1/20) of the cDNA mix was used in 50 µl PCR. A 452 bp fragment of GAPDH was amplified. C-: negative control; C+: positive control; M: 100 bp ladder.

Resources

FAQ

Yes, please follow the Supplementary Protocol 'Purification of cytoplasmic RNA from animal cells using the RNeasy Mini Kit' (RY25).

FAQ ID -1257

The RNA content and RNA make up of a cell depend very much on its developmental stage and the type of cell. To estimate the approximate yield of RNA that can be expected from your starting material, we usually calculate that a typical mammalian cell contains 10–30 pg total RNA.

The majority of RNA molecules are tRNAs and rRNAs. mRNA accounts for only 1–5% of the total cellular RNA although the actual amount depends on the cell type and physiological state. Approximately 360,000 mRNA molecules are present in a single mammalian cell, made up of approximately 12,000 different transcripts with a typical length of around 2 kb. Some mRNAs comprise 3% of the mRNA pool whereas others account for less than 0.1%. These rare or low-abundance mRNAs may have a copy number of only 5–15 molecules per cell.

FAQ ID -2946

Several kit options are available for this application. We recommend using the PAXgene Blood RNA System, which enables the collection, stabilization and transportation of 2.5 ml human whole blood samples, and subsequent rapid and efficient isolation of cellular RNA.

Other products for the isolation of RNA from whole human blood are the QIAamp RNA Blood Mini Kit and the RNeasy Midi Kit for processing up to 1.5 ml and 10 ml human whole blood, respectively.

FAQ ID -304

The exact composition of Buffer RLT is confidential. This buffer is a proprietary component of RNeasy Kits. Buffer RLT contains a high concentration of guanidine isothiocycanate, which supports the binding of RNA to the silica membrane. Buffer RLT can be purchased separately (cat. no. 79216)

Note: note that ß-mercaptoethanol should be added to Buffer RLT before use to effectively inactivate RNAses in the lysate (10 µl ß-Mercaptoethanol per 1 ml Buffer RLT).

FAQ ID -2793

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 ID -290

Yes, please follow the User-Developed Protocol 'Isolation of RNA from leukocytes in milk using QIAGEN RNeasy Kits' (RY12).

FAQ ID -952

Ribonucleases are the principal threat to any RNA isolation procedure. In addition, copurification of inhibitory contaminants is a major problem when isolating RNA from certain tissue sources. To minimize the threat, gloves should be worn at all times, and special care must be taken to use RNase-free reagents and labware.

In addition, tissue/cell lysis steps are typically carried out with lysis buffers containing guanidine isothiocyanate, a potent protein denaturant. It is very important to use a sufficient amount of lysis buffer during RNA isolation. We recommend using at least 10x volume of lysis buffer to tissue/cell pellet.

In general, for fast purification of high-quality RNA we recommend QIAGEN’s RNeasy Kits. It is more challenging to isolate high-quality RNA from tissue samples than from cultured cells, especially those tissues containing high levels of RNase, or difficult-to-homogenize tissues. Examples of such tissues include liver, heart, skin, and conjunctive tissues. Many tissue samples also contain difficult-to-remove contaminants (such as polysaccharides, collagen, fats, lipids or fibrous components) that may interfere with subsequent enzymatic reactions if not removed from the RNA preparation. For purification of high-quality RNA from difficult tissues we recommend QIAGEN’s RNeasy Plus Universal Kit.

FAQ ID -2656

Although it is possible to use the QIAquick Nucleotide Removal Kit and QIAquick PCR Purification Kit for RNA purification, the conditions are not optimized for RNA, nor are RNase-free conditions guaranteed. The RNeasy MinElute Cleanup Kit is recommended for clean-up, concentration and desalting of RNAs above 200 bases in length.

FAQ ID -490

At this time, RNeasy spin columns are not sold separately.

FAQ ID -159

Yes. Even though buffer RDD in the RNase-Free DNase Set is optimized for on-column DNase digestion, the buffer is also well-suited for efficient DNase digestion in solution. Please note that the reaction must be cleaned up after the off-column DNase digest to remove the enzyme and buffer RDD, which will interfere with subsequent RT reactions.

A protocol for in-solution DNase digestion using the RNase-Free DNase Set can be found in Appendix C of the RNeasy MinElute Cleanup Handbook. For subsequent RNA Cleanup, use either the RNeasy MinElute Cleanup Kit, or follow the instructions for RNA Cleanup in the RNeasy Mini Handbook.

FAQ ID -619

The RNeasy Lipid Tissue Mini and Midi Kits are designed to extract RNA from up to 100mg and 500mg of fatty tissue, respectively.

FAQ ID -467

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728

Pancreas is very high in RNases. Therefore, it is important to minimize the time between harvesting the tissue and snap freezing or stabilization in RNAprotect Tissue Reagent. Use of 3-5% ß-mercaptoethanol in Buffer RLT instead of 1% may also improve the results.

FAQ ID -624

The efficiency of downstream applications depends strongly on the purity of the RNA sample used. Pure RNA should yield an A260/A230 ratio of around 2 or slightly above; however, there is no consensus on the acceptable lower limit of this ratio. Possible candidates that can increase the A230 include “salt”, carbohydrates, peptides, and phenol (or aromatic compounds in general). In our experience, the increased absorbance at 230 nm in RNA samples is almost always due to contamination with guanidine thiocyanate, present at very high concentrations in the lysis buffer or extraction reagent used in most RNA purification procedures.

Please find an article discussing the effect of low 260/230 ratios in RNA preparations on downstream applications on page 7 of QIAGEN Newsletter March 15, 2010 . In summary, we found that concentrations of guanidine thiocyanate of up to 100 mM in an RNA sample do not compromise the reliability of downstream applications.

FAQ ID -2248

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12

The exact composition of Buffer RW1 is confidential. Buffer RW1 is a proprietary component of RNeasy Kits. Buffer RW1 contains a guanidine salt, as well as ethanol, and is used as a stringent washing buffer that efficiently removes biomolecules such as carbohydrates, proteins, fatty acids etc., that are non-specifically bound to the silica membrane. At the same time, RNA molecules larger than 200 bases remain bound to the column.

Note: Buffer RW1 should not be used for isolation of small RNAs, for example, microRNAs or fragmented RNA from formalin-fixed tissues, as these smaller fragments will be washed away. Buffer RWT should be used instead.

FAQ ID -2796