RNeasy 96 QIAcube HT Kit
For automated high-throughput isolation of RNA from animal and human cells and tissue
The RNeasy 96 QIAcube HT Kit enables simple, automated purification of total RNA or total RNA, including miRNA, from animal and human cells and tissue samples on the QIAcube HT system. Using proven RNeasy silica-membrane technology in a convenient 96-well format, contaminants and inhibitors are removed to yield high-quality RNA ready for downstream analysis.
The RNeasy 96 QIAcube HT Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Performance
Purification of RNA on QIAcube HT is simple and reliable with the RNeasy 96 QIAcube HT Kit and provides linear sample recovery with no significant well-to-well variation across a 96-well plate (see figure Linear RNA recovery and Reproducible 96-well RNA purification).
This economical, easy-to-use system provides the same results as other trusted QIAGEN purification technologies (see figure Automated purification of RNA from tissues). Analysis of QIAcube HT purified samples shows high RNA Integrity Scores (RIS) when analyzed with the QIAxcel Advanced system, and performance similar to other automated QIAGEN purification solutions (see figure High-quality RNA from tissues). The RNeasy 96 QIAcube HT Kit provides consistent yields from small amounts of starting sample material (see table Typical total RNA yields from cells with the RNeasy 96 QIAcube HT Kit).
Principle
The RNeasy 96 QIAcube HT Kit enables automated purification of total RNA or total RNA, including miRNA, from 5 x 105 animal or human cells or <40 mg tissue using the QIAcube HT instrument. Six instrument protocols are available, depending on the starting material and desired size distribution of purified RNA. Cell samples can be purified directly using the RNeasy 96 QIAcube HT Kit and the dedicated kit protocol. Tissue samples (<40 mg) and samples for purification of small RNA are processed following sample lysis in QIAzol Lysis Reagent. The procedures yield high-quality RNA that performs well in downstream applications.
The RNeasy 96 QIAcube HT Kit combines the selective binding properties of a silica-based membrane with a high-throughput 96-well format, and is designed for fully-automated, simultaneous processing of 24–96 samples on the QIAcube HT instrument.
Procedure
Cell lysis is performed manually under highly denaturing conditions with guanidine thiocyanate to immediately inactivate RNases and ensure purification of intact RNA. After the run is started, ethanol is added to the lysate to provide appropriate binding conditions, and the samples are then applied to the wells of the RNeasy 96 plate. Total RNA binds and contaminants are efficiently washed away. High-quality RNA is then eluted in a small volume of RNase-free water, ready for use in any downstream application.
With the automated procedure for purification of total RNA from animal or human cells, all RNA molecules longer than 200 nucleotides are purified. This provides enrichment for mRNA, since most RNAs <200 nucleotides (such as 5.8S rRNA, 5S rRNA, and tRNA, which together comprise 15–20% of total RNA) are selectively excluded. The size distribution of the purified RNA is comparable to that obtained by centrifugation through a CsCl cushion, where small RNA does not sediment efficiently. For purification of total RNA from tissue, or of total RNA including small RNA, from cells and tissues, we recommend first using QIAzol Lysis Reagent as the lysis buffer. Detailed procedures are provided in the kit handbook.
Applications
The RNeasy 96 QIAcube HT Kit provides efficient, high-throughput RNA sample preparation for research use in fields such as drug screening and basic research. The purified RNA is ready to use in any downstream application, including:
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