DP408-RNAstore Reagent-180123

RNAstore Reagent
For immediate stabilization of the nucleic acid in harvested animal tissues

RNAstore Reagent
Cat. no. 4992727

Storage
RNAstore Reagent should be stored dry at room temperature (15-30°C)
and are stable for 24 months under these conditions. Storage of RNAstore
Reagent at lower temperatures may cause precipitation. Before using the
reagent, redissolve the precipitate by heating to 37°C.

Introduction
RNAstore reagent is a liquid, non-toxic tissue preservation reagent. It can
penetrate into tissue cells rapidly and protect RNA in non-frozen cells in
situby effectively inhibiting RNase activity, making it more suitable for
the analysis of tissue gene expression profile. After harvesting, tissues
are immediately soaked in RNAstore for preservation without causing
RNA degradation, so that the sample does not have to be processed
immediately or frozen in liquid nitrogen. RNAstore Reagent can be widely
used for various animal tissues, including brain, heart, kidney, spleen, liver,
lung and thymus, etc.

Recommended amount of RNAstore Reagent for different tissues (rat):

Important Notes Before Starting

1. RNAstore Reagent is only suitable for fresh animal tissues that have not
   been frozen.
2. To ensure rapid and reliable stabilization of RNA even in the inner parts
   of solid tissues, the sample must be cut into slices less than 0.5 cm thick
   and then stored with 10 volumes of RNAstore Reagent. If the slices are
   thicker than 0.5 cm, the reagent will diffuse too slowly into the interior
   of the sample and RNA degradation will occur.

Protocol

1. Before cutting the tissue sample, estimate the volume of the sample to
   be stabilized in RNAstore Reagent. Add at least 10 volumes of RNAstore
   reagent (For instance: 100 mg tissue with 1 ml RNAstore Reagent)
2. Cut the tissue sample from the animal, and if necessary, cut it into slices
   less than 0.5 cm thick.
   Note: For effective RNA stabilization, the tissue sample must be
   less than 0.5 cm thick and the tissue sample need to be submerged
   completely into RNAstore Reagent.
3. Store the tissue submerged in RNAstore Reagent for up to 4 weeks at
   2-8°C, up to 7 days at 15-30°C, or up to 1 day at 37°C.
   For archival storage at -20°C or -80°C, first incubate the tissue overnight
   in the reagent at 2-8°C. Then remove the tissue from the reagent, and
   transfer it to -20°C or -80°C for storage.
   Note: Lower temperature is recommended for longer storage (e.g.,
   2-8°C for up to 4 weeks instead of 37°C or room temperature; -20°C or
   -80°C for longer storage).
   RNAstore stabilized tissues stored at -20°C or -80°C can be thawed at
   room temperature and frozen again for up to 20 freeze-thaw cycles
   without affecting RNA quality or yield.
   If transporting tissue samples in RNAstore Reagent, ensure that the
   tissues always remain submerged in the reagent.
4. After storage, purify RNA using a TIANGEN RNA isolation product (Trizol,
   RNAprep Pure, RNAsimple).

Other Application

1. RNAstore Reagent could not be used to store leaf samples, since its wax
   epidermis would block the osmosis of RNAstore.
2. For cultured cells, precipitate cells first, then wash the cell pellet with
   PBS, then re-suspend cells with PBS and stored with 2 time volumes
   of RNAstore. Downstream RNA purification can be processed after
   RNAstore removal by centrifugation. There is also another protocol for
   RNA purification experiments without the step of RNAstore removal.
   Centrifugation: Since the medium concentration of RNAstore is higher
   than regular culture medium, so normal speed concentration would
   not able to precipitate cells from RNAstore. Discard RNAstore after
   centrifugation. (HeLa cell need to be centrifuge at around 3000 × g, other
   cells may not able to tolerate this speed, or higher speed is required.)
   Direct extraction: Extract by adding 10 time volumes of RNA extraction
   buffer into cell-RNAstore mix.
3. For bacteria, first precipitate by centrifugation, then wash the pellet
   with PBS, then re-suspend with PBS and stored with 2 time volumes
   of RNAstore. Downstream RNA extraction procedures are same as
   procedures for cultured cells. E. colicould be stored in RNAstore for 1
   month at 4°Cwithout degradation of RNA.
4. For leukocyte, first separate leukocyte from serum and red blood cell,
   then follow the instruction of cultured cells. Do not store RNA from
   whole blood, serum and plasma into RNAstore, because their protein
   concentrations are too high which will form precipitate after the mix
   with RNAstore.

DP408-RNAstore Reagent-180123

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