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RNAlater™ Stabilization Solution (Invitrogen™)

RNAlater® RNA Stabilization Solution stabilizes and protects cellular RNA in intact, unfrozen tissue samples, eliminating the need to immediately process tissue samples or to freeze samples in liquid nitrogen for later processing. Tissue pieces can be harvested and submerged in RNAlater® RNA Stabilization Solution for storage without jeopardizing the quality or quantity of RNA obtained after subsequent RNA isolation. Advantages of using RNAlater® RNA Stabilization Solution:

Effectiveness—stabilize RNA for 1 day at 37°C, 1 week at 25°C, 1 month at 4°C, or indefinitely at -20°C
Simplicity—a single reagent that immediately inactivates RNases and stabilizes RNA within tissues or cells
Convenience—no need to freeze samples in liquid nitrogen or rush samples back to the lab freezer
Mobility—perfect for tissue collection "in the field"
Versatility—compatible with many RNA isolation procedures, including most RNA isolation kits

Applications
RNAlater® RNA Stabilization Solution has been tested on a variety of mammalian tissues, plants, E. coli, Xenopus, fish, and Drosophila. It is ideal for:

• Protecting RNA integrity in tissues rich in RNases
• Collecting samples from different time points without having to process the samples from each time point immediately
• Archiving tissues for future microdissection
• Submerging animal cavities or organs to stabilize RNA during lengthy dissection procedures
• Collecting samples at locations (e.g., hospitals, field sites, the space shuttle) where immediate RNA isolation is not possible
• Shipping samples on wet ice or even at room temperature if shipped overnight

RNAlater® RNA Stabilization Solution procedure
The dissected tissue (less than 0.5 cm in any one dimension) is simply submerged in approximately 5 volumes of RNAlater® solution at room temperature. The solution permeates the cells, stabilizing the RNA. The sample can then be stored indefinitely at -20°C (the tissue does not freeze), at 4°C for up to a month, or at 25°C for up to a week. For RNA isolation, the tissue is simply removed from RNAlater® solution and treated as though it had just been harvested. Most tissues can be transferred directly to a lysis buffer and homogenized. Samples treated with RNAlater® solution and then frozen can be ground with mortar and pestle or thawed and processed like fresh tissue without concern for cell rupture and release of RNases since the RNases have already been inactivated. Cells can be spun out and then added to lysis buffer, or in some cases, RNAlater® solution can be added along with the cells directly to the lysis buffer.

Compatible with a variety of procedures
RNAlater® RNA Stabilization Solution is compatible with one-step RNA isolation methods, such as TRIzol® Reagent; with glass binding methods such as Qiagen's RNeasy™ or the Ambion® RNAqueous® kit; with acid phenol extraction methods such as the Ambion® ToTALLY RNA™ kit; and with methods that use oligo(dT) selection of mRNA, such as the Ambion® Poly(A)Purist™ kit. In-house research, as well recently published independent research, indicates that the use of RNAlater® RNA Stabilization Solution for tissue storage does not affect the outcome of subsequent RNA expression analysis experiments compared to other processing methods.

RNAlater™ Stabilization Solution with Manual (Invitrogen™)

RNAlater® RNA Stabilization Solution stabilizes and protects cellular RNA in intact, unfrozen tissue samples, eliminating the need to immediately process tissue samples or to freeze samples in liquid nitrogen for later processing. Tissue pieces can be harvested and submerged in RNAlater® RNA Stabilization Solution for storage without jeopardizing the quality or quantity of RNA obtained after subsequent RNA isolation. Advantages of using RNAlater® RNA Stabilization Solution:

Effectiveness—stabilize RNA for 1 day at 37°C, 1 week at 25°C, 1 month at 4°C, or indefinitely at -20°C
Simplicity—a single reagent that immediately inactivates RNases and stabilizes RNA within tissues or cells
Convenience—no need to freeze samples in liquid nitrogen or rush samples back to the lab freezer
Mobility—perfect for tissue collection "in the field"
Versatility—compatible with many RNA isolation procedures, including most RNA isolation kits

Applications
RNAlater® RNA Stabilization Solution has been tested on a variety of mammalian tissues, plants, E. coli, Xenopus, fish, and Drosophila. It is ideal for:

• Protecting RNA integrity in tissues rich in RNases
• Collecting samples from different time points without having to process the samples from each time point immediately
• Archiving tissues for future microdissection
• Submerging animal cavities or organs to stabilize RNA during lengthy dissection procedures
• Collecting samples at locations (e.g., hospitals, field sites, the space shuttle) where immediate RNA isolation is not possible
• Shipping samples on wet ice or even at room temperature if shipped overnight

RNAlater® RNA Stabilization Solution procedure
The dissected tissue (less than 0.5 cm in any one dimension) is simply submerged in approximately 5 volumes of RNAlater® solution at room temperature. The solution permeates the cells, stabilizing the RNA. The sample can then be stored indefinitely at -20°C (the tissue does not freeze), at 4°C for up to a month, or at 25°C for up to a week. For RNA isolation, the tissue is simply removed from RNAlater® solution and treated as though it had just been harvested. Most tissues can be transferred directly to a lysis buffer and homogenized. Samples treated with RNAlater® solution and then frozen can be ground with mortar and pestle or thawed and processed like fresh tissue without concern for cell rupture and release of RNases since the RNases have already been inactivated. Cells can be spun out and then added to lysis buffer, or in some cases, RNAlater® solution can be added along with the cells directly to the lysis buffer.

Compatible with a variety of procedures
RNAlater® RNA Stabilization Solution is compatible with one-step RNA isolation methods, such as TRIzol® Reagent; with glass binding methods such as Qiagen's RNeasy™ or the Ambion® RNAqueous® kit; with acid phenol extraction methods such as the Ambion® ToTALLY RNA™ kit; and with methods that use oligo(dT) selection of mRNA, such as the Ambion® Poly(A)Purist™ kit. In-house research, as well recently published independent research, indicates that the use of RNAlater® RNA Stabilization Solution for tissue storage does not affect the outcome of subsequent RNA expression analysis experiments compared to other processing methods.

RNAlater™-ICE Frozen Tissue Transition Solution (Invitrogen™)

A novel reagent for transitioning frozen tissue to a state enabling easy extraction of high-quality RNA; frozen tissues are simply submerged in Ambion® RNAlater®-ICE and allowed to thaw overnight at –20°C. It is provided in one bottle containing 25 mL. Once thawed, the tissues can be processed like fresh tissues using standard RNA isolation procedures. No more laborious grinding of frozen tissue to safeguard the RNA.

• Process previously frozen tissues like freshly harvested samples
• Thawing tissues in RNAlater®-ICE protects RNA from degradation
• No more tissue pulverizing with mortar and pestle and awkward transfer of powder to tube
• Easily apportion frozen tissue samples for multiple experiments

Quick Freezing Tissues Preserves RNA
Often, tissues that need to be stored prior to RNA isolation are "snap" or "flash" frozen on dry ice or in liquid nitrogen to preserve RNA integrity. RNA in tissue is stable while frozen at –80°C, but thawing the tissue for RNA purification can result in RNA degradation. This is true even if the tissue thaws while in the denaturation solution.

Processing Frozen Tissues is Problematic
Frozen tissues are typically ground with a chilled mortar and pestle in order to maintain RNA quality. Liquid nitrogen must be added to the mortar to keep the sample frozen while it is ground. Powdered tissue can also thaw during transfer to a homogenization vessel. This often results in formation of clumps that do not readily disperse in the lysis solution, resulting in RNA degradation and loss.

Process Frozen Tissue Without Jeopardizing RNA Integrity
RNAlater®-ICE solves all of these problems. Simply submerge frozen tissue samples in 10 volumes of the solution and store overnight at –20°C (the solution will remain as liquid at these temperatures). As the tissue thaws, RNA integrity is protected. Once treated, tissue can be safely stored at 4°C or even at room temperature (for a limited period of time) and can be further dissected or processed prior to homogenization in a standard RNA isolation lysis buffer. Thus the same frozen tissue sample can be used multiple times for different experiments without compromising RNA integrity.

THE RNA Storage Solution (Invitrogen™)

THE Ambion® RNA Storage Solution is a buffer that provides greater RNA stability than standard 0.1 mM EDTA or TE Buffer. This 1 mM sodium citrate, pH 6.5 +⁄- 0.1 buffer is provided in 10 tubes containing 1.0 mL each.

The last step in every RNA isolation protocol is to resuspend the purified RNA pellet. After painstakingly isolating the RNA, it is crucial that the pellet be suspended and stored in a safe, RNase-free solution. THE RNA Storage Solution has two features that minimize base hydrolysis of RNA: a low pH, and sodium citrate, which is an efficient chelating agent. THE RNA Storage Solution is compatible with all common RNA applications such as reverse transcription, in vitro transcription, northern analysis, and nuclease protection assays. THE RNA Storage Solution is prepared from the purest reagents available, under scrupulously monitored conditions in a state-of-the-art manufacturing facility. It is then subjected to rigorous quality control procedures to guarantee their purity and performance.