RNeasy Protect Cell Mini Kit

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

* Leukocytes were isolated on a Ficoll gradient. Data kindly provided by Pascale Cornillet-Lefebvre, Hôpital Robert Debré, Reims, France.

Assay result	RNAprotect Cell Reagent	RNAprotect Tissue Reagent
RNA yield using RNeasy technology (µg)	10	3.5
Real-time RT-PCR of abl (CT value)	25	26

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

* 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).

Cell storage conditions	RIN 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

 

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.

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.

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