miRNeasy 96 Advanced QIAcube HT Kit

• Phenol-free miRNA isolation from human, mouse or rat serum and plasma samples
• Simple and reliable automated processing for time and cost savings
• High-quality miRNA ready for use in any downstream application
• Consistent miRNA yields from small amounts of starting material

The miRNeasy 96 Advanced QIAcube HT Kit is designed for automated high-throughput purification of cell-free total RNA – primarily miRNA and other small RNA – from small volumes of serum and plasma on the QIAcube HT. 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 miRNeasy 96 Advanced QIAcube HT Kit is the medium- to high-throughput version of the miRNeasy Serum/Plasma Advanced Kit and provides the same yield and quality of RNA using phenol-free isolation technology (see figures  Performance comparable with manual kit using equal volumes) and  Performance comparable with manual kit using equal amounts of RNA). The miRNeasy 96 Advanced QIAcube HT Kit efficiently isolates total RNA, including miRNA, from human and animal serum and plasma samples from 50 µl to 200 µl ( Processing different amounts of plasma). The kit is compatible with the majority of blood collection devices (see figure  Compatibility with different blood collection tubes). The phenol-free protocol uses easy-to-automate spin column technology in 96-well format and minimize hands on time. Unlike kits from other suppliers, the miRNeasy 96 Advanced QIAcube HT Kit combines ease-of-use and a phenol-free protocol without any compromise on RNA quality or yield (see figure  Comparable or better yields and performance).

The miRNeasy 96 Advanced QIAcube HT Kit enables automated purification of cell-free total RNA – primarily miRNA and other small RNA, from 20–200 µl of human or animal plasma using the QIAcube HT instrument. Selective binding properties of a silica membrane are combined with a high-throughput 96-well format for fully automated, simultaneous processing of 24–96 samples on the QIAcube HT instrument. The kit combines guanidine-based lysis of samples, an inhibitor removal step and silica-membrane-based purification of total RNA.

The system provides two protocols, a transfer protocol to move the lysate from the protein precipitation step to the RNeasy 96 plate and the cell-free RNA isolation protocol to extract miRNA from the lysate. The procedures yield high-quality RNA that performs well in downstream applications .

Lysis is performed manually under highly denaturing conditions with guanidine thiocyanate to immediately inactivate RNases and ensure purification of intact RNA (see figure  miRNeasy 96 Advanced QIAcube HT Kit procedure.)

After the run is started, isopropanol is added to the RNA-containing supernatant to provide appropriate binding conditions for all RNA molecules from approximately 18 nucleotides (nt) upwards. The samples are afterwards transferred (manually or automated) to the wells of the RNeasy 96 plate. Total RNA binds and contaminants are efficiently washed away. High-quality RNA is then eluted in RNase-free water, ready for use in any downstream application.

The miRNeasy 96 Advanced QIAcube HT Kit isolates total cell-free and exosomal RNA from plasma or serum. Downstream applications include:

• Biomarker discovery
• Expression profiling
• Next-generation sequencing
• Quantitative, real-time RT-PCR
• Microarray analysis