RNAlater 说明书

IMPORTANT! Before using this product, read and understand the

“Safety Information” appendix in this document.

■ Product Description
■ Product Information
■ Guidelines for Use of RNAlater® Solution
■ Storage in RNAlater® Solution
■ RNA Isolation from Samples in RNAlater® Solution
■ Quality Control
■ Appendix A Safety Information
■ Documentation and Support
Product Description

RNAlater® Tissue Collection: RNA Stabilization Solution is an

aqueous tissue storage reagent that rapidly permeates most tissues

to stabilize and protect RNA in fresh specimens. It eliminates the

need to immediately process or freeze samples; the specimen can

simply be submerged in RNAlater® Solution and stored for analysis

at a later date.

Samples in RNAlater® Solution can be stored for extended periods

under conditions where RNA degradation would normally take

place rapidly .Tissues can be stored indefinitely in

RNAlater® Solution at –20°C or below.

 

•Store RNAlater® Solution at room temperature.

• If any precipitation of RNAlater® Solution is seen, heat it to

37°C and agitate to redissolve it.

RNAlater® Solution can be used for RNA preservation with most

tissues, cultured cells, bacteria, and yeast. It may not be effective in

tissues that are poorly penetrated by the solution, such as waxy

plant tissue and bone.

RNAlater® Solution has been extensively tested with animal tissues,

including brain, heart, kidney, spleen, liver, testis, skeletal muscle,

fat, lung, and thymus. It has also been proven effective for RNA

preservation in E. coli, Drosophila, tissue culture cells, white blood

cells, and some plant tissues. For more information, go to

www.invitrogen.com/site/us/en/home/support/technical-

support.html.

RNAlater® Solution is compatible with most RNA isolation methods.

Samples stored in RNAlater® Solution have been used successfully

with TRI Reagent® Solution (P/N AM9738), and all of Ambion®

RNA isolation kits and reagents, including: the TōTALLY RNA™ Kit,

the PARIS™ Kit, the mirVana™ miRNA Isolation Kit, and the

RNAqueous® and Poly(A)Purist™ product families.

samples

DNA can be isolated from RNAlater® Solution-stored samples. For

more information, go to www.invitrogen.com/site/us/en/home/

support/technical-support.html.

samples

Proteins are also preserved in RNAlater® Solution. RNAlater®

Solution will denature proteins; therefore, protein obtained from

samples stored in it will be suitable for applications such as Western

blotting or 2D gel electrophoresis, but not for applications that

require native protein.

 

•Use RNAlater® Solution with fresh tissue only; do not freeze

tissues before immersion in RNAlater® Solution.

• Before immersion in RNAlater® Solution, cut large tissue

samples to ≤ 0.5 cm in any single dimension.

• Place the fresh tissue in 5–10 volumes of RNAlater® Solution.

• Most samples in RNAlater® Solution can be stored at room

temperature for 1 week without compromising RNA quality, or

at –20°C or –80°C indefinitely.

• Do not freeze samples in RNAlater® Solution immediately;

store at 4°C overnight (to allow the solution to thoroughly

penetrate the tissue), remove supernatant, then move to –20°C

or –80°C for long-term storage.

Note: We offer RNAlater®-ICE (P/N AM7030) to recover tissues that

have already been frozen. RNAlater®-ICE renders frozen tissues

pliant enough for homogenization while maintaining the low

temperatures needed to protect the RNA from degradation.

 RNAlater® Solution does not disrupt the structure of tissues; thus,

tissue that has been equilibrated in RNAlater® Solution can be

removed from the solution, sectioned into smaller pieces, and

returned to RNAlater® Solution, if desired.

Small organs such as mouse liver, kidney and spleen can be stored

whole in RNAlater® Solution.

Plant Tissue 

Plant tissues that have natural barriers to diffusion, such as waxy

coatings on leaves, will often require disruption to allow RNAlater®

Solution access to the tissue. However, many plant tissues can

simply be submerged in RNAlater® Solution whole; we have

successfully isolated intact RNA from tobacco leaf explants, entire

Arabidopsis and alfalfa seedlings, and from potato shoot tips.

Pellet cells according to the protocols followed by your laboratory.

Remove supernatant and then add 5–10 volumes RNAlater®

Solution. The cells can be washed in PBS before resuspending in

RNAlater® Solution, if desired.

Blood and Plasma 

White blood cells can be effectively preserved in RNAlater® Solution

when separated from the red blood cells and sera and treated as

tissue culture cells. RNAlater® Solution can also be added to small

volumes of anticoagulated whole blood, sera, and plasma; however,

the procedure is not presented here—see the Ambion RiboPure™-

Blood Kit (P/N AM1928) protocol for detailed instructions.

Yeast 

Pellet up to 3 x 108 cells (centrifuge at 12,000 x g for 2 min). Remove

supernatant and immediately resuspend the pellet in 0.5–1 mL of

RNAlater® Solution. Yeast cells can be stored in RNAlater® Solution

for up to 8 hr at 25°C, or up to a week at 4°C.

For long-term storage, incubate the cells in RNAlater® Solution for

1 hr. Repellet the cells (centrifuge at >12,000 x g for 5 min), remove

supernatant, flash freeze, and store at –80°C.

Bacteria  

RNAlater® Solution is bacteriostatic; although bacteria do not grow

in it, the cells remain intact. E. coli stored in RNAlater® Solution for

1 month at 4°C are intact and yield undegraded RNA.

Storage at –80°C

Storage at –80°C is recommended for archival samples and will

provide optimal preservation. Samples can be stored at –80°C

indefinitely. RNAlater® Solution will freeze at –80°C.

To prepare samples for storage at –80°C, first incubate the samples

in RNAlater® Solution overnight at 4°C to allow thorough

penetration of the tissue, then transfer to –80°C. To expedite thawing

of the samples, we recommend removing the tissue, or pelleting

cells, from the RNAlater® Solution before freezing at –80°C.

Samples can subsequently be thawed at room temperature and

refrozen without significantly affecting the amount or the integrity

of the recoverable RNA.

Storage at –20°C

Storage at –20°C can also be used for archival samples. Samples will

not freeze at –20°C, but crystals may form; this will not affect

subsequent RNA isolation. Samples can be stored at –20°C

indefinitely.

To prepare samples for storage at –20°C, first incubate the samples

in RNAlater® Solution overnight at 4°C to allow thorough

penetration of the tissue, then transfer to –20°C.

Samples can subsequently be thawed at room temperature and

refrozen without affecting the amount or the integrity of the

recoverable RNA.

Storage at 4°C

Most samples can be stored in RNAlater® Solution at 4°C for up to

1 month without significant RNA degradation.

Place samples in the coolest environment available. If ambient

temperature is above 25°C, incubate the samples in RNAlater®

Solution on ice for a few hours, if possible, before storing at ambient

temperature.

Storage at 25°C (room temperature)

Most samples can be stored at 25°C in RNAlater® Solution for up to

1 week without significant loss of RNA quality. After 2 weeks at

25°C, RNA generally appears slightly degraded (marginally

acceptable for Northern analysis, but still of sufficient quality for

nuclease protection assays or RT-PCR analysis).

Storage at 37°C

RNA isolated from samples stored at 37°C is intact after a 24 hour

incubation, but is partially degraded after 3 days.

RNase inactivation is reversible; do not rinse RNAlater® Solution

from samples before using. Blot tissues with a wipe, or pellet cells to

remove excess RNAlater® Solution.

Tissue

Retrieve tissue from RNAlater® Solution with sterile forceps, quickly

blot away excess RNAlater® Solution with an absorbent lab wipe or

paper towel, and then submerge the sample in RNA isolation lysis

solution. Homogenize tissue promptly after placing it in lysis/

denaturation solution.

Cells

There are two options for isolating RNA from cells stored in

RNAlater® Solution. The preferred method is to remove the solution

from the cells prior to extraction. Alternatively, cells in RNAlater®

Solution can be used directly for RNA extraction. Because of the

greater volume that the cells are in, this method generally requires

additional lysis solution.

• Removal of RNAlater® Solution prior to extraction

Because of the density of RNAlater® Solution, greater

centrifugal forces are required to pellet cells from RNAlater®

Solution than from normal media. Generally, cells become

much less fragile when stored in RNAlater® Solution and can be

centrifuged at high speed without lysis. Most cell types can be

centrifuged at 5000 x g without damage to the cells. Since

different cell types vary in their ability to withstand centrifugal

forces, we recommend testing the centrifugal speed with an

expendable sample. Alternatively, dilute the RNAlater®

Solution by adding an equal volume of ice cold PBS (or other

buffered solution) immediately before centrifugation to reduce

the density of the solution, then centrifuge at normal speeds.

• RNA extraction from cells in RNAlater® Solution

One-step phenol-based disruption/extraction solutions, such as

Ambion TRI Reagent® Solution or RNAwiz™ Reagent

(available only in Japan), can be used to purify RNA from cells

suspended in RNAlater® Solution. This can be done by adding

ten volumes of the one-step solution to the cell mixture, and

proceeding normally. When RNAwiz Reagent is used in this

way, it may be necessary to dilute the aqueous phase before the

RNA precipitation step. See below for more information.

Lysates from RNAlater® Solution-treated samples often require more

force to pass through glass-fiber filters than lysates from untreated

samples. Therefore, it may be necessary to use centrifugation instead

of vacuum pressure to pass lysates through glass-fiber filters.

When using one-step RNA isolation products such as TRI Reagent

Solution or RNAWIZ Reagent (available only in Japan), on RNAlater®

Solution-preserved samples, the aqueous phase will occasionally

appear cloudy; this will not adversely affect RNA recovery or

quality.

With RNAWIZ Reagent, there may be a problem getting the aqueous

phase to mix with isopropanol at the precipitation step because of

RNAlater® Solution carryover. If this occurs, add a mixture of

50% water, 50% isopropanol until the solution becomes clear and the

two phases mix. The amount of water/isopropanol required will

depend on how much RNAlater® Solution was carried over; if the

sample was mostly RNAlater® Solution, as much as an equal volume

may be needed.

RNAlater® Solution undergoes quality assurance testing to verify

that its composition is invariant from lot to lot.

 

WARNING! GENERAL SAFETY. Using this product in a

manner not specified in the user documentation may result in

personal injury or damage to the instrument or device. Ensure

that anyone using this product has received instructions in

general safety practices for laboratories and the safety

information provided in this document.

• Before using an instrument or device, read and understand

the safety information provided in the user documentation

provided by the manufacturer of the instrument or device.

• Before handling chemicals, read and understand all

applicable Safety Data Sheets (SDSs), and use appropriate

personal protective equipment (gloves, gowns, eye

protection, etc.). To obtain SDSs, see the “Documentation

and Support” section in this document.

Chemical safety WARNING! GENERAL CHEMICAL HANDLING. To

minimize hazards, ensure laboratory personnel read and

practice the general safety guidelines for chemical usage,

storage, and waste provided below, and consult the relevant

SDS for specific precautions and instructions:

• Read and understand the Safety Data Sheets (SDSs) provided

by the chemical manufacturer before you store, handle, or work

with any chemicals or hazardous materials. To obtain SDSs, see

the “Documentation and Support” section in this document.

• Minimize contact with chemicals. Wear appropriate personal

protective equipment when handling chemicals (for example,

safety glasses, gloves, or protective clothing).

• Minimize the inhalation of chemicals. Do not leave chemical

containers open. Use only with adequate ventilation (for

example, fume hood).

• Check regularly for chemical leaks or spills. If a leak or spill

occurs, follow the manufacturer's cleanup procedures as

recommended in the SDS.

• Handle chemical wastes in a fume hood.

• Ensure use of primary and secondary waste containers. (A

primary waste container holds the immediate waste. A

secondary container contains spills or leaks from the primary

container. Both containers must be compatible with the waste

material and meet federal, state, and local requirements for

container storage.)

• After emptying a waste container, seal it with the cap provided.

• Characterize (by analysis if necessary) the waste generated by

the particular applications, reagents, and substrates used in

your laboratory.

• Ensure that the waste is stored, transferred, transported, and

disposed of according to all local, state/provincial, and/or

national regulations.

IMPORTANT! Radioactive or biohazardous materials may

require special handling, and disposal limitations may apply.

WARNING! Depending on the samples used on the

instrument, the surface may be considered a biohazard. Use

appropriate decontamination methods when working with

biohazards

WARNING! BIOHAZARD. Biological samples such as

tissues, body fluids, infectious agents, and blood of humans

and other animals have the potential to transmit infectious

diseases. Follow all applicable local, state/provincial, and/or

national regulations. Wear appropriate protective equipment,

which includes but is not limited to: protective eyewear, face

shield, clothing/lab coat, and gloves. All work should be

conducted in properly equipped facilities using the

appropriate safety equipment (for example, physical

containment devices). Individuals should be trained

according to applicable regulatory and company/institution

requirements before working with potentially infectious

materials. Read and follow the applicable guidelines and/or

regulatory requirements in the following:

• U.S. Department of Health and Human Services guidelines

published in Biosafety in Microbiological and Biomedical

Laboratories found at: www.cdc.gov/biosafety

• Occupational Safety and Health Standards, Bloodborne

Pathogens (29 CFR§1910.1030; www.access.gpo.gov/nara/

cfr/waisidx_01/29cfr1910a_01.html).

• Your company’s/institution’s Biosafety Program protocols

for working with/handling potentially infectious

materials.

• Additional information about biohazard guidelines is

available at: www.cdc.gov

 

• www.invitrogen.com/sds

or

• www.appliedbiosystems.com/sds

Note: For the SDSs of chemicals not distributed by Life

Technologies, contact the chemical manufacturer.

• www.invitrogen.com or

• www.appliedbiosystems.com

At the website, you can:

• Access worldwide telephone and fax numbers to contact

• Search through frequently asked questions (FAQs)

• Submit a question directly to Technical Support

• Search for user documents, SDSs, vector maps and sequences,

application notes, formulations, handbooks, certificates of

analysis, citations, and other product support documents

• Obtain information about customer training

• Download software updates and patches