miRNeasy 96 Kit
For 96-well purification of microRNA and total RNA from tissues and cells
The miRNeasy 96 Kit enables purification of total RNA, which includes RNA from approximately 18 nucleotides (nt) upwards from all types of animal tissues and cells, including difficult-to-lyse tissues. Alternatively, an miRNA-enriched fraction and a total RNA (>200 nt) fraction can be purified separately (for separate purification, an additional RNeasy 96 plate is required; for economical purchase of an additional RNeasy 96 plate and wash buffers, we recommend ordering an RNeasy 96 Kit). Manual purification is carried out using the Centrifuge 4K15C or the Centrifuge 4-16K and, optionally, the QIAvac 96 vacuum manifold.
Purification of total RNA including miRNA allows direct comparison of miRNA expression levels with those of housekeeping reference genes or any other mRNA of interest. miRNeasy Kits enable efficient enrichment of RNA down to approximately 18 nt in size. The presence of very small RNAs is clearly visible after purification using an miRNeasy Mini Kit, in contrast to samples prepared using an alternative kit (see figure "Effective enrichment of small RNA"). miRNeasy Kits efficiently purify RNA from both tissues and cells, even when low amounts of starting material are used (see figure "Effective purification from a range of starting amounts").
RNA can be purified with the kit from a variety of samples, including difficult-to-lyse tissues (see figure "Efficient copurification from a wide range of tissues"). miRNeasy procedures eliminate the possibility of contamination with salts or phenol which could interfere with later analyses (see figure "Highly pure RNA without phenol carryover"). In addition to higher purity, miRNeasy Kits offer superior yields to alternative methods of miRNA purification, such as using TRIzol Reagent (see figure "miRNeasy Kit outperforms TRIzol"). RNA prepared using miRNeasy Kits is highly pure and ready for use in sensitive downstream applications.
The miRNeasy 96 Kit combines phenol/guanidine-based sample lysis and silica membrane-based RNA purification with the speed of vacuum and/or spin processing. QIAzol Lysis Reagent, included in the kit, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.
The miRNeasy 96 Kit enables purification of total RNA which includes RNA from 18 nucleotides (nt) upwards. Alternatively, an miRNA-enriched fraction and a total RNA (>200 nt) fraction can be can be purified separately (see flowchart "miRNeasy procedures"). The separate miRNA-enriched fraction may be required to reduce background in particularly sensitive downstream applications. For this specialized protocol, enabling enrichment of miRNAs and other small RNAs (<200 nt), 2 RNeasy 96 plates are required for every 96 samples.
The miRNeasy 96 Kit allows purification of miRNA with total RNA for use in a variety of applications, including:
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