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RNeasy Protect Cell Mini Kit

For stabilization and purification of total RNA from cells
  • Immediate RNA stabilization in sorted or cultured cells
  • No need to remove medium prior to cell stabilization
  • Storage and shipping of cells without dry ice
  • Archiving of cells at –20ºC or –80ºC
  • Purification of high-quality RNA in minutes

The RNeasy Protect Cell Mini Kit includes RNAprotect Cell Reagent, which stabilizes RNA and DNA in sorted or cultured cells and allows convenient and reliable transport of the cells at ambient temperature. The stabilized cells can be stored or shipped at 30°C (1 day), 15–25°C (7 days), or 2–8°C (4 weeks). The RNeasy Protect Cell Mini Kit provides the RNeasy Plus Mini Kit for purification of total RNA. Stabilized cells are lysed and spun through gDNA Eliminator spin columns to effectively remove genomic DNA. High-quality RNA is then purified using RNeasy spin columns. The purification procedure can be automated on the QIAcube.

Cat No./ID: 74624
RNeasy Protect Cell Mini Kit (50)
$429.00
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RNAprotect Cell Reagent (50 ml) and RNeasy Plus Mini Kit (50)
The RNeasy Protect Cell Mini 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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Effective inhibition of PMA induction.

Incubation of PMA-stimulated Jurkat cells in RNAprotect Cell Reagent prevents PMA induction of c-fos expression and prevents degradation of c-fos transcript, demonstrating stabilization of RNA by RNAprotect Cell Reagent (each bar represents the mean of duplicate cell samples).

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RNeasy Protect Cell procedure.
Performance

The RNeasy Protect Cell Mini Kit is ideal for gene expression analysis experiments where cells need to be delivered from one lab to another (see Table 1).

Table 1. Higher RNA yields from leukocytes* preserved for 10 days in RNAprotect Cell Reagent
Assay resultRNAprotect Cell Reagent RNAlater Reagent
RNA yield using RNeasy technology (µg) 10 3.5
Real-time RT-PCR of abl (CT value) 25 26
* Leukocytes were isolated on a Ficoll gradient. Data kindly provided by Pascale Cornillet-Lefebvre, Hôpital Robert Debré, Reims, France.

Cells in RNAprotect Cell Reagent can be stored or transported for up to 1 day at 30ºC, up to 7 days at room temperature (15–25ºC), or up to 4 weeks at 2–8ºC, and can also be stored long-term at –20ºC or –80ºC (see Table 2, and figures "Effective inhibition of PMA induction" and "More reliable quantification of CD83 transcript").

Table 2. Intact RNA from Jurkat cells stored at ambient temperature
Cell storage conditionsRIN value of purified RNA*
30ºC (1 day) 9.8
25ºC (7 days) 9.7
2–8ºC (28 days) 10
–20ºC (28 days) 9.8
–80ºC (28 days) 9.8
* RNA integrity is assessed on the Agilent 2100 Bioanalyzer, which gives RIN values on a scale of 1 (completely degraded RNA) to 10 (completely intact RNA).

Principle

The RNeasy Protect Cell Mini Kit provides a complete RNA protection and isolation solution, from sample harvesting to pure RNA, in one kit. Proven RNeasy technology, combined with the RNA-stabilizing properties of RNAprotect Cell Reagent, allows purification of high-quality, intact RNA. Sorted or cultured cells are mixed with RNAprotect Cell Reagent, which immediately stabilizes the cellular RNA, ensuring reliable results in downstream gene expression analysis. RNAprotect Cell Reagent can be added directly to cells in culture medium or buffer, eliminating the need to wash or process cells (e.g., by trypsin treatment). Alternatively, RNAprotect Cell Reagent can be added to pelleted cells. Following stabilization, RNeasy technology simplifies total RNA isolation by combining the stringency of guanidine-isothiocyanate lysis with the speed and purity of silica-membrane purification.

Procedure
Sorted or cultured cells are mixed with RNAprotect Cell Reagent, which immediately stabilizes the cellular RNA. After RNA stabilization, total RNA is purified using the RNeasy Plus Mini Kit (see flowchart "RNeasy Protect Cell procedure"). The cells in RNAprotect Cell Reagent are centrifuged, and the resulting cell pellet is lysed and homogenized in Buffer RLT Plus. The lysate is then passed through a gDNA Eliminator spin column, which rapidly and effectively removes genomic DNA. Ethanol is added to the lysate, which is then applied to an RNeasy spin column. After centrifugation, total RNA binds to the membrane of the RNeasy spin column. Contaminants are efficiently washed away, and high-quality total RNA is eluted in 30–50 µl of RNase-free water. Purification of DNA, or of RNA and DNA, requires the additional purchase of a QIAGEN DNA purification kit. Details are provided in the RNAprotect Cell Reagent Handbook.
Applications

Cells in RNAprotect Cell Reagent can be transported at ambient temperature, avoiding the need to freeze cell pellets in liquid nitrogen and to ship the frozen pellets on dry ice. RNA purified using the RNeasy Plus Mini Kit is ideally suited for downstream applications that are sensitive to low amounts of DNA contamination, such as:

  • Quantitative real-time RT-PCR
Features
Specifications
Applications PCR, qPCR, real-time RT-PCR, microarray
Elution volume 30–50 µl
Format Spin column
Main sample type Cultured cells
Processing Manual (centrifugation)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Total RNA
Sample amount Max. 10e7 cells
Stabilization Yes
Technology Silica technology
Yield Varies

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Kit Handbooks (2)
RNAprotect Cell Reagent ─ 培養細胞中のRNAを迅速に安定化; RNeasy Protect Cell Mini Kit ─ 培養細胞中のトータルRNAの迅速な安定化およびトータルRNA精製  
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RNAprotect Cell Reagent
For immediate stabilization of nucleic acids in cells

RNeasy Protect Cell Mini Kit
For immediate stabilization of nucleic acids in cells
and subsequent total RNA purification
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