miRNeasy Tissue/Cells Advanced Mini Kit
For phenol-free purification of microRNA and total RNA from tissues and cells
The miRNeasy Tissue/Cells Advanced Mini Kit is the second generation of the well-established miRNeasy Mini Kit. This improved version combines a phenol-free advanced chemistry with gDNA Eliminator Columns for safe and convenient RNA isolation. miRNeasy Tissue/Cells Advanced Mini Kit enables purification of total RNA, including small RNAs ≥18 nucleotides, from animal tissues and cells. The kit uses spin columns for low-throughput RNA purification. Furthermore, the entire procedure can be automated on QIAcube Connect.
The miRNeasy Tissue/Cells Advanced Mini Kit is a phenol-free alternative to the miRNeasy Mini Kit with similar RNA yields and quality (see figure "High recovery of miRNA without the need for phenol"), and enables efficient enrichment of RNA down to approximately 18 nucleotides in size. The miRNeasy kits efficiently purify RNA from up to 30 mg frozen tissue or 1 x 107 cells, but also works with lower amounts of starting material. The miRNeasy Tissue/Cells Advanced Mini Kit includes the gDNA Eliminator procedure allowing high, reproducible RNA yields and efficient genomic DNA elimination for sensitive applications (see figure "Total RNA extraction: effective genomic DNA removal"). Total RNA with Agilent RIN values close to 10 is routinely obtained from tissues and cultured cells. Unlike kits from other suppliers, the miRNeasy Tissue/Cells Advanced Mini Kit combines ease-of-use and a phenol-free protocol without any compromise on RNA quality or yield (see figures "Competitive performance" and "Fast procedure").
The miRNeasy Tissue/Cells Advanced Mini Kit combines a guanidine-based sample lysis with an inhibitor removal step and a silica-membrane-based purification of total RNA, which includes miRNA and other small RNA. Buffer RLT, included in the kit, contains guanidine thiocyanate which facilitates lysis and denaturation of RNases and other proteins. Therefore, RNA in samples lysed in buffer RLT are stable and protected from degradation.
Sample lysis is achieved by adding Buffer RLT to tissue or cell samples followed by a thorough disruption and homogenization step to ensure complete lysis and inactivation of RNases. Next, Buffer AL is added to the lysate to optimize gDNA removal by the gDNA Eliminator Spin Column. Depending on the type and amount of starting material, an optional on column DNase digestion can be carried out (not included in the kit). Buffer RPP is added to the flow-through to precipitate inhibitors (mostly proteins that are highly concentrated in tissue and cell samples and might interfere with the RNA isolation or the downstream analysis) by centrifugation. The supernatant containing the total RNA is transferred into a new reaction tube, and isopropanol is added to provide appropriate binding conditions for all RNA molecules ≥18 nucleotides. The sample is then applied to a RNeasy Mini Spin Column, where the total RNA binds to the membrane and all contaminants are efficiently washed away. High-quality RNA, including miRNA and other small RNA, are eluted in a small volume of RNase-Free Water.
The miRNeasy Tissue/Cells Advanced Mini Kit allows purification of miRNA and total RNA for a variety of applications, including:
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