For 96-well purification of total RNA from cells
  • High-throughput RNA purification
  • Fast and convenient sample processing
  • Reproducible yields from 10 to 500,000 cells
  • High-quality RNA for any application
  • No organic extraction or precipitation

The RNeasy 96 Kit provides high-throughput purification of total RNA from up to 96 cultured-cell samples using silica-membrane RNeasy 96 plates. Manual purification is carried out using the QIAGEN 96-Well-Plate Centrifugation System and/or the QIAvac 96 vacuum manifold. Automated purification using the related RNeasy 96 BioRobot 8000 Kit is performed on the BioRobot Universal System, which processes up to 96 or 192 samples per run followed by real-time RT-PCR setup.
For optimal results it is recommended to use this product together with QIAvac 96.

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RNeasy 96 Kit (12)
For 12 x 96 total and cytoplasmic RNA preps: 12 RNeasy 96 Plates, Elution Microtubes CL, Caps, S-Blocks, AirPore Tape Sheets, RNase-Free Reagents and Buffers
74182
$8,238.00
RNeasy 96 Kit (4)
For 4 x 96 total and cytoplasmic RNA preps: 4 RNeasy 96 Plates, Elution Microtubes CL, Caps, S-Blocks, AirPore Tape Sheets, RNase-Free Reagents and Buffers
74181
$3,002.00
The RNeasy 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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High-quality RNA for sensitive analysis of a low-copy transcript.|Reproducible yields of high-quality RNA.|RT-PCR of RNA from as few as 100 cells.|Repeatability of fully automated RNA purification.|RNeasy 96-well plate.|
RNA was isolated from 1 to 1 x 105 HeLa S3 cells using the RNeasy 96 BioRobot 8000 procedure. Total RNA was eluted in 100 µl RNase-free water, and 5 µl was used for RT-PCR. Quantitative, real-time one-step RT-PCR analysis was carried out on an ABI Sequence Detection System using the QuantiTect Probe RT-PCR Kit with primers and probe specific for the low-copy c-fos transcript. [A] Amplification plot. [B] CT values. Error bars represent standard deviation from 8 different samples for each cell number.|[A] Agarose gel analysis. HeLa cells (5 x 105) from a homogeneous cell culture were pelleted in each well of a 96-well cell-culture plate and the total RNA isolated using the RNeasy 96 system. Half of each eluate was analyzed on a formaldehyde agarose gel. [B] The corresponding northern blot, hybridized with a 32P-labeled GAPDH probe. [C] Analysis of RNA yields. Each square represents the yield from a preparation originating from one well of the 96-well cell-culture plate.|RT-PCR of cytoplasmic RNA isolated with the RNeasy 96 Kit from the indicated numbers of HeLa cells. 10 µl (1/10) of the eluate was reverse-transcribed with oligo-dT primer, and 5 µl (1/5) of the cDNA mix was used in a 100 µl PCR in which a 214 bp fragment of beta-actin was amplified. C-: no template; C+: beta-actin in vitro transcript; M: 100 bp DNA ladder.|RNA was isolated from 96 aliquots (5 x 104 cells each) of a HeLa S3 cell culture using the RNeasy 96 BioRobot 8000 procedure. Quantitative, real-time one-step RT-PCR was set up in the same protocol on the BioRobot 8000 workstation, using the QuantiTect Probe RT-PCR Kit with primers and dual-labeled probe specific for the low-copy c-myc transcript. Threshold cycles (CT) are shown for all 96 samples. The mean CT was 21.34 ± 0.34 (mean ± standard deviation), representing a CV of 1.6%.|The RNeasy 96 plate contained in the RNeasy 96 Kit.|
Performance
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").
Principle

The RNeasy 96 system provides a standardized, fast, reliable method for isolation of high-quality RNA from large numbers of samples (see table). The simple procedure is the key to the speed and efficiency of RNeasy 96 technology. 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 (see figure "RNeasy 96-well 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. Since the RNeasy procedure enriches for RNA species >200 nt, RNA yield does not include 5S rRNA, tRNAs, or other low-molecular-weight RNAs.

Specifications of the RNeasy 96 Kit
SpecificationDetails
Format RNeasy 96-well plates with 1.2 ml collection microtubes
Sample source Animal or human cells, in vitro transcripts, or enzymatic reactions
Sample size 10–5 x 105 cells
Binding capacity Up to 100 µg RNA per well
Yield* from 1 x 105 cells 

HeLa: 1.6 µg
LMH: 1.3 µg
COS-7: 3.1 µg
Huh7: 2.0 µg
Jurkat: 1.4 µg

Elution volume 45–140 µl
* Amounts can vary due to developmental stage, growth conditions used, etc.

Procedure

The key to the speed and efficiency of RNeasy 96 technology is the simple 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 (see figure "RNeasy 96-well 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.

RNA isolation can also be automated on the BioRobot 9604 workstation using the RNeasy 96 BioRobot 9604 Kit. Fully automated RNA isolation is possible on the BioRobot 8000 workstation or BioRobot Universal System using the RNeasy 96 BioRobot 8000 Kit.

Applications

The RNeasy 96 Kit consistently provides the highest-quality RNA for:

  • High-throughput RNA applications in gene expression analysis
  • Sensitive applications such as quantitative, real-time RT-PCR
  • Pharmacological and toxicological research
Feature
Specifications
Applications PCR, qPCR, real-time PCR
Elution volume 45–140 µl
Format 96-well plate
Main sample type Cultured cells
Processing Manual (centrifugation or vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 5 x 10e5
Technology Silica technology
Time per run or per prep 30–40 minutes
Yield 1.3–3.1 µg

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Kit Handbooks
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User-Developed Protocols
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One milliliter of milk contains approximately 50,000-200,000 leukocytes (with an average of approximately 100,000 leukocytes). In a BVDV-infected animal up to 1-30% of the leukocytes may be infected with the virus. Each infected leukocyte typically contains 10-10,000 BVDV entities.
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