RNAprotect Reagents and Tubes

For immediate stabilization of RNA in tissues, cultured cells and bacterial cultures

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

RNAprotect Tissue Reagent (50 ml)

Cat. No. / ID: 76104

50 ml RNAprotect Tissue Reagent for stabilization of RNA in 25 x 200 mg tissue samples

ReagentTube

RNAprotect Reagent

RNAprotect Tubes

Sample type

Tissue

Bacteria

Cells

Size

RNAprotect Tissue Reagent (50 ml)

RNAprotect Tissue Reagent (250 ml)

Inquire

RNAprotect Reagents and Tubes are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Immediate stabilization of RNA and RNA protection
Convenient and safe handling at room temperature
Easy collection and storage of tissues in reclosable tubes
Stabilization of Gram-positive and Gram-negative bacteria
No need to remove medium from sorted or cultured cells

Product Details

RNAprotect Reagents and Tubes provide immediate RNase inactivation and stabilization of RNA in tissues, cultures of Gram-positive and Gram-negative bacteria and sorted or cultured cells to preserve the gene expression profile and enable reproducible purification of high-quality RNA. Samples are stabilized at room temperature prior to RNA isolation procedures.

Performance

RNAprotect Tissue Reagent ensures immediate RNase inactivation and stabilizes and protects the RNA expression pattern of samples during harvest and storage (see figure " RNAprotect Reagent prevents degradation of mRNA in tissues"). Immediate protection with RNAprotect technology ensures that downstream analyses truly reflect the expression profile of the intact tissues. Samples can be archived without risk of RNA degradation, even after multiple freeze–thaw cycles (see figures “ Stable RNA in tissues: different temperatures and freeze–thaw cycles” and “ RNAprotect Tissue Tubes prevent RNA changes in tissues”).

RNAprotect Cell Reagent prevents induction of gene expression in treated cells and preserves and stabilizes transcript levels during storage of cell samples (see figures " Effective inhibition of PMA induction of c-fos expression" and " More reliable quantification of CD83 transcript").

RNAprotect Bacteria Reagent is suitable for use with a wide range of bacterial species, both Gram positive (e.g., Staphylococcus aureus and Mycobacterium avium) and Gram negative (e.g., Escherichia coli and Salmonella typhimurium) and prevents RNA changes (see figures “ RNAprotect Bacteria Reagent prevents mRNA degradation” and “ GeneChip analysis”). Bacteria can be grown in either minimal or complex medium prior to isolation of RNA.

GeneChip analysis.
RNeasy purified RNA from RNAprotect stabilized samples stored under a wide variety of conditions is ideal for downstream processes such as northern blotting and RT-PCR.

See figures

RNAprotect Reagent prevents degradation of mRNA in tissues.

Stable RNA in tissues: different temperatures and freeze–thaw cycles.

RNAprotect Tissue Tubes prevent RNA changes in tissues.

Effective inhibition of PMA induction.

More reliable quantification of CD83 transcript.

RNAprotect Bacteria Reagent prevents mRNA degradation.

GeneChip analysis.

Principle

Once a biological sample is harvested, its RNA becomes extremely unstable. RNAprotect Reagents and Tubes enable immediate RNase inactivation, stabilization of the RNA and preservation of the RNA expression pattern. Gene induction or down-regulation triggered by sample manipulation is also prevented, allowing accurate analysis of gene expression. This is particularly important for bacterial expression profiles. Enzymatic degradation of RNA leads to reduction or loss of many transcripts because bacterial mRNA molecules have very short half lives of only a few minutes.

RNAprotect Tissue Reagent quickly permeates tissues resulting in immediate stabilization of the RNA. RNAprotect Cell Reagent provides immediate stabilization of RNA in sorted or cultured cells. Cells are stabilized at room temperature and can be stored or transported at ambient temperature prior to RNA purification.

Procedure

For RNA protection and sample storage, tissue samples are submerged in an appropriate volume of RNAprotect Tissue Reagent immediately after harvesting. RNAprotect Tissue Tubes provide premeasured volumes of RNAprotect Tissue Reagent in reclosable tubes for convenient handling and sample storage. The reagent preserves RNA for up to 1 day at 37°C, 7 days at 18–25°C, or 4 weeks at 2–8°C, allowing processing, transportation, storage, and shipping of samples without liquid nitrogen or dry ice. Alternatively, the samples can also be placed at –20°C or –80°C for archival storage.

RNAprotect Cell Reagent immediately stabilizes the gene expression profile in cells. With other technologies, it is necessary to wash or process the cells (e.g., trypsin treatment) prior to adding an RNA stabilization reagent. Changes in expression that may occur during the washing or processing steps are eliminated by adding RNAprotect Cell Reagent directly to cells in culture medium or buffer. Instantaneous RNA stabilization with RNAprotect Cell Reagent can also be extended to pelleted cells not in medium or storage solution. The amount of starting material must not exceed 1 x 10⁷ cells.

For bacterial samples, two volumes of RNAprotect Bacteria Reagent are added directly to 1 volume of bacterial culture prior to RNA isolation, providing immediate stabilization of RNA. The stabilization allows time for efficient bacterial lysis using a choice of protocols: enzymatic lysis, mechanical disruption, or a combination of both methods. QIAGEN recommends the TissueLyser for efficient mechanical disruption.

Applications

RNA stabilized in RNAprotect Reagents and Tubes can be used for:

Total RNA purification
miRNA purification
miRNA purification

Supporting data and figures

RNAprotect Reagent prevents degradation of mRNA in tissues.

Resources

RNAprotect Tissue Tubes - For collection of harvested animal tissues with immediate stabilization of the gene expression profile, and subsequent transport and storage; RNAprotect Tissue Reagent - For immediate stabilization of the gene expression profile in harvested animal tissues

Download Safety Data Sheets for QIAGEN product components.

FAQ

Yes, we recommend to use the RNeasy Protect Cell Mini Kit which contains RNAprotect Cell Reagent, the first RNA stabilization reagent for cultured cells in the market optimized for a broad range of cell types and cell culture formats.

FAQ ID -1215

Bacterial cultures stabilized in RNAprotect Bacteria Reagent can be stored at room temperature for up to 3 hours. Alternatively, pellets obtained after centrifugation of the bacterial culture/RNAprotect Bacteria Reagent mixture can be frozen and stored at -20ºC for up to 2 weeks, or at -70ºC for up to 4 weeks.

FAQ ID -614

Yes, RNAprotect Bacteria Reagent and the RNeasy Mini Kit are compatible. You can also purchase QIAGEN's RNeasy Protect Bacteria Kits, which will contain RNAprotect Bacteria Reagent and RNeasy chemistry. Protocols for isolating RNA from bacteria after using the Reagent for bacterial RNA stabilization can be found in the RNAprotect Bacteria Reagent Handbook.

FAQ ID -797

The RNeasy 96 Universal Tissue Kit enables high-throughput purification of RNA from any animal or human tissue sample, including difficult-to-lyse fibrous and fatty tissues. For single tube format RNA purification, the RNeasy Plus Universal Kit is also available.

Please refer to the Selection guide for RNA isolation for all sample types to find the optimal solution for your sample source.

FAQ ID -627

Yes, please follow the protocol in the RNAprotect Bacteria Reagent Handbook in Appendix D: Stabilization and Purification of Bacterial RNA Using RNAprotect Bacteria Reagent, the RNeasy Lipid Tissue Mini Kit, and the TissueLyser.

FAQ ID -1020

RNAprotect Tissue Reagent is recommended for stabilization and protection of RNA in animal tissues. It is not recommended for cultured cells because it may be difficult to pellet and remove them from the reagent prior to RNA isolation (see FAQ 364).

Instead, we recommend to use the RNAprotect Cell Reagent. This reagent is for immediate RNA stabilization in sorted or cultured cells with no need to remove medium

FAQ ID -941

For Laser Captured Microdissection, its best to either flash-freeze the tissue and do cryosectioning, or to use FFPE material.

FAQ ID -608

In general, this is possible and has been done successfully in our labs with brain tissue. However, if the starting material for RNA isolation is extremely fatty, it will be difficult for the aqueous RNAprotect Tissue Reagent to permeate into the tissue and therefore, to sufficiently stabilize it.

FAQ ID -856

Yes, RNAprotect Cell Reagent of the RNeasy Protect Cell Mini Kit can be purchased separately in a bulk size of 250 ml for processing large cell culture volumes.

FAQ ID -1217

The RNAprotect Tissue Reagent penetrates the sample by diffusion and protects RNA immediately upon contact with the surface layer and outer portions of solid tissues. In order to ensure reliable stabilization of RNA even in the inner parts of solid tissues, we recommend incubation for at least 45 minutes in the reagent prior to RNA isolation using RNeasy Kits. Note that tissue size is critical for successful RNA stabilization with RNAprotect technology. Samples must be cut into slices less than 0.5 cm thick for rapid and reliable stabilization in interior tissue parts. RNA degradation will occur in tissue slices that are too thick.

For archival storage in the freezer, first incubate the sample overnight in the reagent at 2–8°C. Please see detailed instructions for storage under step 5 of the “Protocol: Stabilization of RNA in Harvested Animal Tissues” in the RNAprotect Handbook.

FAQ ID -647

No, we do not recommend to use the RNAprotect Tissue Reagent to stabilize flash-frozen tissue. It should only be used to stabilize fresh tissue.

FAQ ID -757

Yes, please follow the User-Developed Protocol 'Preparation of RNAprotect preserved tissues for pathological studies' (PROM-15329).

FAQ ID -950

We do not recommend to store previously isolated RNA in RNAprotect Tissue Reagent, since it will be very difficult to recover isolated RNA from it. Purified RNA can be stored at −30 to −15°C or −90 to −65°C in sterile RNase-free water. Under these conditions, we found no detectable RNA degradation after 1 year of storage.

FAQ ID -337

The RNA content and make up of a bacterial cell is highly dependent on the type of bacteria, and the developmental and physiological state of the cell. To estimate the approximate yield that can be expected from your starting material, we usually calculate that a typical bacterial cell contains 100 fg of total RNA.

FAQ ID -2949

Different samples of varying water content may affect the point at which the reagent will form a solid at low temperature. Other factors can be temperature variation from freezer to freezer, or even within the same freezer (e.g., different temperatures near the door and in the back region of the freezer). Regardless, the performance of the RNAprotect Tissue Reagent and the RNA quality are not affected by freezing of the reagent.

FAQ ID -758

Yes, but you have to make sure that the RNAprotect Bacteria Reagent is diluted in PBS or water prior to use (2 volumes reagent : 1 volume PBS or water) in order to achieve its optimal working concentration. The diluted reagent is directly added to the bacteria growing on the solid substrate, and reagent and bacteria are mixed thoroughly. The reagent is not recommended for stab cultures, since sufficient access of the reagent to all bacteria for immediate RNA stabilization cannot be guaranteed.

FAQ ID -615

For RNA isolated on the BioRobot EZ1 and BioRobot M48:

The RNA can be directly applied to the Agilent Bioanalyzer, since it is being denatured in the final protocol steps of these isolation procedures.

For RNA prepared with all other QIAGEN RNA Isolation Products:

We recommend to denature the samples in a water bath for 2 min at 70°C, and then place them directly on ice prior to loading them onto the Agilent Bioanalyzer.

FAQ ID -528

If the cells in RNAprotect Tissue Reagent cannot be collected by centrifugation, please try one of the following suggestions:

1) Add 600 µl Buffer RLT to a maximum of 200 µl sample volume, and proceed with step 3 of the "RNeasy Mini Protocol for Isolation of Total RNA from Animal Cells" in the RNeasy Mini Handbook. Load the lysate onto the column in successive aliquots in step 5 of the protocol.

2) If cells are floating on the surface of the RNAprotect Tissue Reagent, try removing the reagent by pipetting from underneath. Leave behind approximately 100 ul of RNAprotect Tissue Reagent, and add 350 ul Buffer RLT before proceeding with the protocol "RNeasy Mini Protocol for Isolation of Total RNA from Animal Cells". For every 100 ul of cells in RNAprotect Tissue Reagent, use 250 ul of 96-100% ethanol instead of the 70% ethanol listed in step 4 of the standard protocol.

3) Dilute the sample 10x by adding cold PBS. Pellet cells by centrifugation. Caution: Cells might lyse.

4) Try to pipette the floating cells off the surface.

Note that the above steps are suggestions, rather than official protocol recommendations. Please try a "pilot" run on a test sample first.

For RNA stabilization of cells, we recommend using the RNAprotect Cell Reagent.

FAQ ID -364

Tissues preserved in RNAprotect Tissue Reagent can be stored as long as listed below:

Temp [°C]	Duration
37 °C	1 day
room temp.	1 week
2-8 °C	1 month
-30 to -15 °C	archival
-90 to -65 °C	archival

FAQ ID-3750

We never have seen it in-house for mouse or rat pancreas, but if a lot of fatty tissue is present in the tissue sample it may float on top. Our R&D department has obtained good results with RNAprotect Tissue Reagent preserved mouse and rat pancreas.

FAQ ID -536

We do not recommend to use cells stabilized with RNAprotect Cell Reagent for cell sorting as cell morphology will be altered after adding the reagent, and stabilized cells may lyse during the sorting process.

FAQ ID -1216