Cat. No. / ID: 74634
The QIAwave RNA Plus Mini Kit is a more eco-friendly version of our standard RNeasy Plus Mini Kit, combining high-quality RNA purification with effective gDNA removal using unique gDNA Eliminator columns. Total RNA of high purity and yield is obtained from a wide range of cultured cells and easy-to-lyse tissues.
The QIAwave RNA Plus Mini Kit uses up to 47% less plastic and up to 42% 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 RNA Plus Mini Kit may look different, it’s as easy to use as the RNeasy Plus Mini Kit, and the chemistry and performance are identical.
Please be aware that you will need sterile glass bottles to store the reconstituted buffers.
The QIAwave RNA Plus Mini Kit gives you highly reproducible yields of total RNA from cells and tissues. The performance between our QIAwave RNA Plus Mini Kit and the RNeasy Plus Mini Kit is identical because the chemistry is the same. Further, the kit performs similarly or better compared to a competitor kit (see figure “ RNA yields obtained with QIAwave RNA Plus Mini Kit”). Total RNA isolated with the QIAwave RNA Plus Mini Kit results in similar downstream RT-qPCR performance as total RNA isolated with the RNeasy Plus Mini Kit, as well as similar or better performance compared to a competitor kit. (see figure: “ RT-qPCR Performance of total RNA isolated with QIAwave RNA Plus Mini Kit”)
We have also compared RNA yields obtained with the QIAwave RNA Plus Mini Kit (50) preparing the buffer either by pouring or pipetting and with the RNeasy Plus Mini Kit (50) using standard buffers. All methods result in comparable RNA yields as shown in the figure “ Handling of buffer concentrates”.
The QIAwave RNA Plus Mini procedure integrates QIAGEN’s innovative technology for selective binding of double-stranded DNA with well-established RNeasy technology. Efficient purification of high-quality RNA is guaranteed, without the need for additional DNase digestion. The purified RNA is ready to use and is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as quantitative, real-time RT-PCR and dPCR.
Cells and easy-to-lyse tissues are first lysed and homogenized in highly denaturing guanidine-isothiocyanate–containing Buffer RLT Plus, which immediately inactivates RNases to ensure isolation of intact RNA. The lysate is then passed through a gDNA Eliminator spin column that, in combination with the high-salt buffer, selectively and efficiently removes genomic DNA. Ethanol is added to provide appropriate binding conditions for RNA, and the sample is applied to an RNeasy spin column. These specialized columns contain a silica membrane that specifically binds RNA from lysed cells.
The QIAwave RNA Plus Mini Kit isolates total RNA from up to 107 cells or 30 mg tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 minutes (see flowchart “ QIAwave RNA Plus Mini procedure”). Samples are first lysed and homogenized. The lysate is passed through a gDNA Eliminator spin column, ethanol is added to the flow-through, and the sample is applied to an RNeasy spin column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in 30 µL water.
Different protocols are available for different starting materials. The protocols differ mainly in the lysis and homogenization of the sample. Once the sample is applied to the gDNA Eliminator spin column, the protocols are similar. The procedure provides an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded.
When disrupting and homogenizing tissues in Buffer RLT Plus (supplied with the RNeasy Plus Mini Kit), excessive foaming may occur. This foaming is substantially reduced by adding Reagent DX (supplied separately) to Buffer RLT Plus before starting disruption and homogenization. Reagent DX has been carefully tested with the kit, and has no effect on RNA purity or on downstream applications.
The QIAwave RNA Plus Mini Kit can be automated on the QIAcube Connect using the RNeasy Plus Mini Kit protocol.
RNA purified using the QIAwave RNA Plus Mini Kit is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as RNA-seq and quantitative real-time RT-PCR. The purified RNA can also be used in other applications.
Total RNA isolated from 5x105 Jurkat cells with the QIAwave RNA Plus Mini Kit and the RNeasy Plus Mini Kit was quantified via UV/VIS spectroscopy.
|Applications||RNA-seq, quantitative real-time RT-PCR, end-point RT-PCR, Northern dot and slot blotting, microarray analysis|
|Elution volume||30-50 µL|
|Integrated removal of genomic DNA||Yes|
|Sample type||Cells, tissues|
|Sample amount||107 cells or 30 mg tissue|
|Processing||Manual or automated on QIAcube Connect|
|Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein||Total RNA|
|Time per run||25 minutes|