75144-RNeasy Midi Kit (50)- Qiagen

50 RNeasy Midi Spin Columns, Collection Tubes (15 ml), RNase-free Reagents and Buffers

RNeasy Kits are the Gold standard for total RNA isolation. They provide fast purification of high-quality RNA from small to large amounts of cells, tissues, and yeast using silica-membrane RNeasy spin columns or 96-well plates. Tissue samples can be conveniently stabilized using RNAprotect Tissue Reagent or Allprotect Tissue Reagent, and efficiently disrupted using a TissueRuptor II or TissueLyser II or LT system. The RNeasy 96 Kit enables high-throughput purification of total RNA from up to 96 cultured-cell samples using silica-membrane RNeasy 96 plates. A dedicated RNeasy QIAcube Kit enables automated purification of 1–12 samples on the QIAcube Connect.

Performance

RNeasy Kits deliver highly reproducible yields of total RNA from small to large samples. Total RNA can reliably be purified from small numbers of cells, including a single cell, as well as from small amounts of standard tissues (see figures “Reliable RNA isolation from a single cell”, “Highly reproducible yields for sensitive applications” and “High-quality total RNA from fine needle aspirates”) using the RNeasy Micro Kit. RNA purified using the RNeasy Micro Kit provides maximum sensitivity with precious samples in quantitative gene expression analyses, such as real-time RT-PCR, by efficient on-column digestion of genomic DNA (see figure “Efficient on-column removal of genomic DNA”).
Total RNA is easily purified from animal or human cells, animal or human tissues, and yeast (see table “Total RNA yields obtained with RNeasy Kits” and figure “RT-PCR of RNA from as few as 100 cells”).

The dedicated RNeasy Mini QIAcube Kit, including rotor adapters preloaded with RNeasy spin columns and elution tubes, delivers greater convenience and time savings (see figure “Significant time savings with dedicated QIAcube Kits”).

Total RNA purified with the RNeasy Maxi Kit is of high quality and is suitable for many downstream applications (see figure “High-quality RNA from a variety of samples”). Total RNA is easily purified with the RNeasy Maxi Kit from large amounts of starting material including animal or human cells, animal or human tissues, and yeast cells (see table “Total RNA yields obtained with RNeasy Kits”).

The RNeasy 96 system provides fast and reproducible total RNA purification for high-throughput gene expression profiling. The RNA is suitable for sensitive applications such as quantitative, real-time RT-PCR and microarray analysis (see figure “High-quality RNA for sensitive analysis of a low-copy transcript”). Sample sizes range from 10 to 5 x 105 cells (see figure “RT-PCR of RNA from as few as 100 cells”), and high-quality RNA can be purified from large numbers of samples (see figure “Reproducible yields of high-quality RNA”). Individual variations are low throughout the entire purification process; TaqMan® threshold-cycle values are easily achieved at the end of the process with a coefficient of variation (CV) less than 3% using the QuantiTect Probe RT-PCR Kit (see figure “Repeatability of fully automated RNA purification”).

Procedure

RNeasy technology simplifies total RNA isolation (see table “Amount of starting material for RNeasy procedures”). Samples are first lysed and then homogenized. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the RNeasy silica membrane and RNA binds to the silica membrane, and all contaminants are efficiently washed away. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using a convenient on-column DNase treatment. Pure, concentrated RNA is eluted in water. A variety of special application protocols is also available.

Automated processing on the QIAcube Connect

The QIAcube Connect uses advanced technology to process QIAGEN spin columns, enabling seamless integration of automated, low-throughput sample prep into laboratory workflows. All steps in the purification procedure are fully automated — and up to 12 samples can be processed per run. The QIAcube Connect together with the dedicated RNeasy Mini QIAcube Kit provides fast, easy, and convenient RNA purification.

High-throughput processing in 96-well format

The RNeasy 96 system provides a fast and efficient procedure. Cells can be grown and directly lysed in 96-well cell-culture dishes. After transfer of the lysates to the wells of the RNeasy 96 plate, RNA binds to the silica membrane in each well, and contaminants are washed away. Pure RNA is then eluted in RNase-free water into individual collection tubes suitable for long-term storage and is ready to use for any experiment. The RNeasy 96 Kit includes protocols for total RNA isolation, cytoplasmic RNA isolation, and RNA cleanup. RNeasy 96 plates can be processed on the QIAvac 96 vacuum manifold and/or the 96-Well-Plate Centrifuge System.

Applications

RNA purified with RNeasy technology has A260/280 ratios of 1.9–2.1 (measured in 10 mM Tris·Cl, pH 7.5) and is ideal for use in all applications. Downstream applications include:

• RNA-seq
• Quantitative, real-time RT-PCR
• Real-time RT-PCR starting with as little as one cell
• End-point RT-PCR
• Northern, dot, and slot blotting
• Array analysis
• Poly A+ RNA selection