Cat. No. / ID: 74534
The QIAwave RNA Mini Kit is an eco-friendlier version of our standard RNeasy Mini Kit. The kit uses up to 60% less plastic and up to 57% 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 up to 90% 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:
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 III or LT system.
The QIAwave RNA Mini Kit can be automated on the QIAcube Connect using the RNeasy Mini Kit protocols.
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
|LMH||1 x 106||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||-|
|Liver||10 mg||40 µg||-|
|Lung||10 mg||10 µg||-|
|Spleen||10 mg||35 µg||-|
|S. cerevisiae||1 x 107||25 µg||-|
*Amounts can vary due to developmental stage, growth conditions used, etc.
We have also compared RNA yields obtained with the QIAwave RNA Mini (50) buffer, prepared by pouring or pipetting, and the RNeasy Mini Kit (50) standard buffers. Both methods result in comparable RNA yields as shown in the figure “ Handling buffer concentrates.”
The QIAwave RNA Mini Kit allows you to efficiently purify total RNA from samples such as:
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.
With the QIAwave Kit, you can purify RNA in four simple steps (see flowchart " QIAwave RNA Mini Procedure").
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|
The QIAwave RNA Mini Kit can be automated on the QIAcube Connect using the RNeasy Mini Kit protocols.
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:
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.
Yes, please follow the Supplementary Protocol 'Purification of cytoplasmic RNA from animal cells using the RNeasy Mini Kit' (RY25).
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.
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.
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).
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.
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.
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.).
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.
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.
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.
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.