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RNeasy 96 BioRobot 8000 Kit

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

Cat No./ID: 967152
RNeasy 96 BioRobot 8000 Kit (12)
$6,690.00
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For 12 x 96 total RNA preps on the BioRobot 8000 or BioRobot Universal System: 12 RNeasy 96 Plates, Elution Microtubes CL, Caps, S-Blocks, RNase-Free Reagents and Buffers
The RNeasy 96 BioRobot 8000 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.
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.
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Reproducible yields of high-quality RNA.
[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.
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RT-PCR of RNA from as few as 100 cells.
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.
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Repeatability of fully automated RNA purification.
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%.
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RNeasy 96-well plate.
The RNeasy 96 plate contained in the RNeasy 96 BioRobot 8000 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 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 BioRobot 8000 Kit
Specification Details
Format RNeasy 96-well plates with 1.2 ml collection microtubes
Sample source Animal or human cells
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

Fully automated total RNA isolation using the RNeasy 96 BioRobot 8000 Kit is performed on the BioRobot 8000 workstation or BioRobot Universal System. The BioRobot worktable has the capacity to purify RNA from up to 192 samples in a single run. With the one-plate protocol, the system can carry out both RNA purification and reaction setup in a single run — from living cell cultures in 96-well plates to RT-PCR mixtures, ready to use in real-time gene expression analysis. Filter tips prevent cross-contamination, and precision robotic handling provides minimal well-to-well variation and high repeatability. RNA purification can also be performed manually on the QIAvac 96 vacuum manifold and/or the 96-Well-Plate Centrifugation System using the RNeasy 96 Kit.

Applications

The RNeasy 96 BioRobot 8000 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
Features
Specifications
Applications PCR, real-time PCR, microarray
Elution volume 45–140 µl
For automated processing BioRobot 8000 Workstation or BioRobot Universal System
Format 96-well plate
Main sample type Cultured cells
Processing Automated
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein RNA
Sample amount 5 x 10e5
Technology Silica technology
Yield 1.3–3.1 µg

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Kit Handbooks (1)
For high-throughput RNA purification from animal and human cells, automated on the BioRobot Universal System, BioRobot Gene Expression — Real-Time RT-PCR, or BioRobot 8000
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