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RNeasy Plus 96 Kit

For 96-well purification of total RNA from cells using gDNA Eliminator plates
  • Integrated removal of genomic DNA
  • No need for DNase digestion
  • Fast, convenient sample processing
  • Reproducible RNA yields from up to 2,000,000 cells
  • Highly standardized procedure
The RNeasy Plus 96 Kit provides high-throughput purification of total RNA from cultured cells grown in 96-well plates. The kit combines the use of novel gDNA Eliminator 96 plates with RNeasy 96 plates to ensure purification of RNA that is virtually free of genomic DNA contamination. The fast, standardized purification procedure delivers reproducible yields of high-quality RNA, ensuring reliable results in real-time RT-PCR, microarray analysis, and other downstream applications.
For optimal results it is recommended to use this product together with QIAvac 96 .
Cat No./ID: 74192
RNeasy Plus 96 Kit (12)
€5,542.00
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For 12 x 96 total RNA preps: 12 RNeasy 96 Plates, 12 gDNA Eliminator 96 Plates, Elution Microtubes CL, Caps, S-Blocks, AirPore Tape Sheets, RNase-Free Reagents and Buffers
The RNeasy Plus 96 Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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RNeasy Plus 96 procedure.
1
Effective genomic DNA removal.
Total RNA was purified from Jurkat cells (105, 104, 103, and 102) using the RNeasy Plus 96 Kit. Real-time RT-PCR analysis of the β-actin transcript was performed using a primer–probe set that could amplify and detect both mRNA and genomic DNA sequences. The flat amplification plot for the no RT control (–RT) indicates the absence of genomic DNA.
2
High quality RNA.

Total RNA was purified from the indicated cell lines using the RNeasy Plus 96 Kit and analyzed on the Agilent 2100 Bioanalyzer. The RIN values were close to 10, indicating the high integrity of the purified RNA.

3
Reliable RNA purification.
Total RNA was purified from 10-fold serial dilutions of Jurkat cells (106 to 102 cells) using the RNeasy Plus 96 Kit. Real-time RT-PCR analysis of the β-actin transcript was performed. The linear graph of CT values against cell dilution indicates reliable RNA purification over a wide range of amounts of starting material (R2 = 0.9985).
Performance

The RNeasy Plus 96 Kit is compatible with a wide range of cell lines, providing purification of high-quality RNA with Agilent RIN values of close to 10 (see figure "High-quality RNA"). Complete genomic DNA removal by the kit (see figure "Effective genomic DNA removal") enables specific transcript detection in highly sensitive applications, such as quantitative, real-time RT-PCR analysis of low-abundance targets. In addition, reliable RNA purification from 102 to 106 cells allows real-time RT-PCR quantification over a wide dynamic range (see figure "Reliable RNA purification").

Principle

Cells are first lysed and homogenized in a highly denaturing guanidine-isothiocyanate–containing buffer, which immediately inactivates RNases to ensure isolation of intact RNA. The lysates are then passed through a gDNA Eliminator 96 plate. This 96-well plate, in combination with the high-salt buffer, selectively and efficiently removes genomic DNA. Ethanol is added to provide appropriate binding conditions for RNA, and the samples are applied to an RNeasy 96 plate. Each well of these specially designed 96-well plates contains a silica membrane that specifically binds RNA from lysed cells.

Procedure
High-throughput RNA purification is achieved through the use of RNeasy 96 plates. Prior to application to theRNeasy 96 plate, cell lysates are first passed through a gDNA Eliminator 96 plate, (see figure "RNeasy Plus 96 procedure"). The gDNA Eliminator 96 plates are conveniently processed using a centrifuge (Centrifuge 4-16). RNA purification using RNeasy 96 plates is manual, and comprises 3 simple steps: bind, wash, and elute. The plates are rapidly and conveniently processed using either a centrifuge (Centrifuge 4-16 and Plate Rotor 2 x 96) or a combination of vacuum (QIAvac 96) and centrifuge. Up to 2 x 106 cells can be used as starting material when processing plates using a centrifuge, while up to 1 x 106 cells can be used when processing plates using a vacuum and centrifuge. The procedure provides an enrichment for mRNA since most RNAs <200 nucleotides (such as rRNAs and tRNAs) are excluded. The RNeasy Plus procedure can be modified to allow the purification of total RNA containing small RNAs, such as miRNA, from cultured cells.
Applications

The purified total RNA, which represents the transcriptome state of the cell, is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as Quantitative real-time RT-PCR. The purified RNA can also be used in other applications.

The ease with which RNA is purified in 96-well format makes the kit well suited for high-throughput gene expression analysis in areas such as:
Drug discovery
Biomedical research
Features
Specifications
Applications End-point RT-PCR, quantitative, real-time RT-PCR
Elution volume 45–140 µl
Format 96-well plate
Main sample type Cultured cells
Processing Manual (centrifugation, vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 10 to 5 x 10e5 cells
Technology Silica technology
Yield 1.3–3.1 µg (from 1 x 10e5 cells)

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Kit Handbooks (2)
For purification of total RNA from animal and human cells in 96-well format using gDNA Eliminator 96 plates
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gDNA Eliminator 96 Plate を用いて96ウェルフォーマットにより
動物およびヒト細胞からトータルRNAを精製
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Safety Data Sheets (1)
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