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查看更多产品信息 RNAlater™ Stabilization Solution - FAQs (AM7023M)
42 个常见问题解答
将溶液加热至37°C，保持15分钟，并搅拌溶解。
可使用RNAlater-ICE冻存组织转变溶液浸泡冻存组织，并在–20°C或更低温度解冻过夜。解冻后，可作为新鲜组织进行标准RNA分离处理。
虽然我们还没有进行内部测试，但有证据表明可以使用在RNAlater稳定溶液中保存后的组织进行激光捕获显微切割。请参阅下面的参考文献:
Thelen P, Burfeind P, Grzmil M et al. (2004) cDNA microarray analysis with amplified RNA after isolation of intact cellular RNA from neoplastic and non-neoplastic prostate tissue separated by laser microdissections. Int J Oncol 24(5):1085–1092.
RNA储存在RNAlater溶液中时，在37°C下可保存1天，25°C下可保存1周，4°C下可保存1个月，在-20°C下可长期保存。
可以，使用RNAlater溶液处理后的细胞可通过荧光激活细胞分选法（FACS）进行分析。如果RNAlater溶液的粘性带来问题，则可能需要使用无核酸酶的冷水将其稀释。这不会影响对RNA的保护作用。应尽可能快速并在低温下处理样品。点击此处（https://www.thermofisher.com/us/en/home/references/ambion-tech-support/rna-isolation/tech-notes/facs-into-rnalater-solution-for-gene-profiling.html），阅读关于这一步的更多信息和文献。使用RNAlater溶液处理后的样品也可进行质谱分析。请注意，在去吸附/离子化步骤中，盐可在蛋白质上形成加合物。为避免出现这个问题，可透析样品以除去所有的盐。
不能，RNAlater溶液中的组织必须在4°C保存过夜。
可以，RNAlater溶液已被用于表皮葡萄球菌分离株。下面这篇文献描述了研究人员如何将该溶液用于革兰氏阳性菌：
Handke LD, Conlon KM, Slater SR et al. (2004) Genetic and phenotypic analysis of biofilm phenotypic variation in multiple Staphylococcus epidermidis isolates. J Med Microbiol 53(Pt5):367–344.
RNAlater溶液是抑菌剂。虽然细菌在RNAlater溶液中不会生长，但细胞仍保持完整。将大肠杆菌细胞储存在RNAlater溶液中，在4°C下保存1个月，细胞可保持完整，提取的RNA未降解。
虽然我们尚未对此进行检测，但以下参考文献认为使用RNAlater溶液处理不会使病毒灭活：
•Uhlenhaut C, Kracht M (2005) Viral infectivity is maintained by an RNA protection buffer. J Virol Methods 128(1–2):189–191.
•Kurth A (2007) Possible biohazard risk from infectious tissue and culture cells preserved with RNAlater. Clin Chem3(7):1389–1390.
通过对在RNAlater中-20°C储存2年以上的组织提取的RNA进行评估，已证明在RNAlater溶液中储存不会带来系统性偏差。想要了解更多信息，敬请查看我们的网页（https://www.thermofisher.com/us/en/home/references/ambion-tech-support/rna-isolation/tech-notes/rna-remains-stable-during-long-term-tissue-storage.html）。
从在RNAlater溶液中储存过的组织中可以提取出DNA。请点击此处（https://www.thermofisher.com/us/en/home/references/protocols/nucleic-acid-purification-and-analysis/rna-protocol/genomic-dna-preparation-from-rnalater-preserved-tissues.html）查看实验方案。蛋白质可以在RNAlater溶液中保留；但是，该溶液会使蛋白质变性。因此，从在RNAlater溶液中储存过的组织中提取出的蛋白质仍然适用于免疫印迹法或2D凝胶电泳等应用，但不适用于需要非变性蛋白质的应用。
对于组织样本，我们推荐将组织在4°C下于过量的RNAlater溶液中孵育过夜后取出，使用灭菌的Kimwipe纸擦干，再进行裂解。
对于细胞样本，首先将细胞离心，倒出多余的RNAlater上清液。如果可能，应从细胞沉淀中吸出RNAlater溶液，然后再裂解。在RNAlater溶液中储存过的细胞通常更强韧，使用更高转速离心（5,000 x g）不会导致细胞裂解。
将组织切成任一方向小于0.5 cm的薄片，并浸没于约5倍体积的RNAlater溶液中（如0.5 g样本需要约2.5 mL的RNAlater溶液）。小的器官，如大鼠肾、肝和脾，可整个保存在RNAlater溶液中。细胞样品可以在少量PBS中重悬细胞沉淀，然后加入5-10体积的RNAlater溶液。样品在4°C下可保存1个月，25°C下可保存1周，-20°C下可无限期保存。您可将使用RNAlater溶液处理的组织保存在-20°C下作为存档。
用于RNA提取时，只需将组织从RNAlater溶液中取出，将其作为刚收集的组织进行处理。大多数组织可在裂解缓冲液中直接匀浆处理，但较硬的组织（如骨骼）则需要在液氮中冷冻后再研磨。
只要在进行RNA分离前将RNAlater稳定溶液去除，RNAlater稳定溶液与标准RNA分离试剂盒是兼容的。
RNAlater溶液是一种水溶性组织储存试剂，可在完整、未冻存组织样品中保护细胞RNA。使用RNAlater溶液后不再需要立即处理组织样本，也无需将样本在液氮中冷冻待后续处理。
Heat the solution to 37 degrees C for 15 minutes and agitate to redissolve it.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
RNAlater-ICE Frozen Tissue Transition Solution can be used to submerge a frozen sample, then thaw it overnight at -20 degrees C or colder. Once thawed, tissues can then be processed like fresh tissues using standard RNA isolation procedures.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
While we have not tested this in-house, there is evidence that it is possible to use tissues preserved in RNAlater Stabilization Solution and then perform laser capture microdissection. Please see the reference below:
Thelen P, Burfeind P, Grzmil M et al. (2004) cDNA microarray analysis with amplified RNA after isolation of intact cellular RNA from neoplastic and non-neoplastic prostate tissue separated by laser microdissections. Int J Oncol 24(5):1085-1092.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
RNA stored in RNAlater Stabilization Solution is stable for 1 day at 37 degrees C, 1 week at 25 degrees C, 1 month at 4 degrees C, or long-term at -20 degrees C.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Yes, you should be able to sort cells by fluorescence-activated cell sorting (FACS) after treatment with RNAlater Stabilization Solution. If you run into problems due to the viscosity of RNAlater Stabilization Solution, you may need to dilute it with cold nuclease-free water. This will not affect the protection of the RNA. Samples should be processed quickly and as cold as possible. Read more about this procedure and view references here (http://www.thermofisher.com/us/en/home/references/Invitrogen-tech-support/rna-isolation/tech-notes/facs-into-rnalater-solution-for-gene-profiling.html). You should also be able to perform mass spectrophotometry on samples treated with RNAlater Stabilization Solution. Please note that salt can form adducts on the protein at the desorption/ionization step. To avoid this problem, dialyze the sample to get rid of all salt.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
No, the tissue in RNAlater Stabilization Solution must be stored at 4 degrees C overnight.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Yes, RNAlater Stabilization Solution has been used with Staphylococcus epidermidis isolates. Here's a reference describing how researchers used the solution with gram-positive bacteria:
Handke LD, Conlon KM, Slater SR et al. (2004) Genetic and phenotypic analysis of biofilm phenotypic variation in multiple Staphylococcus epidermidis isolates. J Med Microbiol 53(Pt5):367-344.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
RNAlater Stabilization Solution is bacteriostatic. Although bacteria do not grow in RNAlater solution, the cells remain intact. E. coli cells stored in RNAlater Stabilization Solution for 1 month at 4 degrees C are intact and yield undegraded RNA.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
While we have not tested this in house, these references suggest that the virus remains active after treatment with RNAlater Stabilization Solution:
- Uhlenhaut C, Kracht M (2005) Viral infectivity is maintained by an RNA protection buffer. J Virol Methods 128(1-2):189-191.
- Kurth A (2007) Possible biohazard risk from infectious tissue and culture cells preserved with RNAlater. Clin Chem3(7):1389-1390.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
RNA from RNAlater treated tissue was evaluated after -20 degrees C storage for over 2 years, indicating no systematic bias introduced by storage in RNAlater Stabilization Solution. For more information, visit our webpage at http://www.thermofisher.com/us/en/home/references/ambion-tech-support/rna-isolation/tech-notes/rna-remains-stable-during-long-term-tissue-storage.html.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Yes, DNA can be isolated from samples stored in RNAlater Stabilization Solution. Please view the protocol here: https://www.thermofisher.com/us/en/home/references/protocols/nucleic-acid-purification-and-analysis/rna-protocol/genomic-dna-preparation-from-rnalater-preserved-tissues.html. Proteins will be preserved in RNAlater Stabilization Solution; however, the solution will denature the proteins. Therefore, the protein obtained from samples stored in it will still be suitable for applications such as western blotting or 2D gel electrophoresis, but not for applications that require native protein.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
For your tissue samples, we recommend removing the tissue treated with RNAlater Stabilization Solution from the excess RNAlater solution after incubation overnight at 4 degrees C, blotting the tissue on an absorbent lab wipe or paper towel, and proceeding to lysis.
For your cell samples, pellet the cells, then pour off the excess RNAlater supernatant. Cells are generally less fragile after being stored in RNAlater Stabilization Solution and most cells can be centrifuged at higher speeds without causing cell lysis (5,000 x g).
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Trim the tissue to be less than 0.5 cm in at least one dimension and simply submerge it in 5 volumes of RNAlater Stabilization Solution (e.g., a 0.5 g sample requires about 2.5 mL of RNAlater Stabilization Solution). You can store most samples at 4 degrees C for up to one month, at 25 degrees C for up to one week, or at -20 degrees C indefinitely.
For RNA isolation, simply remove the tissue from RNAlater Stabilization Solution and treat it as though it was just harvested. Most tissues can be homogenized directly in lysis buffer, although harder tissues such as bone may require freezing in liquid nitrogen and grinding.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
RNAlater Stabilization Solution is compatible with standard RNA isolation kits as long as the RNAlater Stabilization Solution is removed prior to the RNA isolation procedure.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
RNAlater Stabilization Solution is an aqueous tissue storage reagent that protects cellular RNA in intact, unfrozen tissue samples. RNAlater Stabilization Solution eliminates the need to immediately process your tissue samples or to freeze them in liquid nitrogen for later processing.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
For the Cells-to-CT lysis reaction, you can use fresh cells, frozen cells, or cells that have been stabilized with RNAlater solution. You just want to ensure that cells were washed once with PBS before going into the lysis reaction.
Yes, for the purpose of RNA preservation in tissue, RNAlater Stabilization Solution will work as well as snap freezing (
https://www.thermofisher.com/us/en/home/references/ambion-tech-support/rna-isolation/tech-notes/rna-remains-stable-during-long-term-tissue-storage.html).
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
We have only tested RNAlater Stabilization Solution for preservation of tissue up to 1 month at 4 degrees C before isolation of RNA. Therefore, we cannot recommend using it for long-term preservation of tissue with no freezing
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Yes, RNAlater Stabilization Solution will freeze at -80 degrees C. We recommend treating your tissue overnight at 4 degrees C with RNAlater and then removing RNAlater prior to freezing at -80 degrees C
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
No, RNAlater Stabilization Solution does not involve chemical fixation or crosslinking. It works by denaturing proteins, including RNase.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Dilute the solution 1:1 with cold PBS and centrifuge at up to 5000 x g (start at lower speed and increase until the cells pellet). Transfer the supernatant to a new tube and discard the pellet.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Yes, you can isolate genomic DNA from tissue preserved in RNAlater Stabilization Solution (Cat. No. AM7024).
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Unfortunately, we do not offer a sterile version of RNAlater Stabilization Solution.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
RNAlater should not impact downstream PCR amplification as long as the sample has been properly cleaned before proceeding with DNA isolation, as stated in the RNAlater Stabilization Solution manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/7020M.pdf) on page 7.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
RNAlater 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. In other words, it is an RNA preserving reagent.
RNaseOUT Recombinant Ribonuclease Inhibitor is a potent non-competitive inhibitor of pancreatic-type ribonucleases such as RNase A, and is used to avoid RNA degradation in a variety of applications such as cDNA synthesis, RT-PCR, and in vitro transcription and translation. RNAseOUT inhibits RNase A, RNAse B, and RNAse C. In other words, RNaseOUT Recombinant Ribonuclease Inhibitor is mainly used in an assay to prevent RNA degradation.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Yes, MagMAX-96 for Microarrays Total RNA Isolation Kit is compatible with the RNAlater solution, because the kit uses Tri-Reagent. Therefore, most of the salts are removed during the lysis steps and partitioned from aqueous RNA layer on top. Additionally, salt carry-over from the RNAlater solution is not an issue.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Both Tempus Blood RNA Tubes and RNAlater Stabilization Solution employ the same effective stabilization of mRNA expression profiles, and eliminate the need to isolate the RNA immediately after sample collection. However, there are some key differences:
- Components in each stabilization solution are different. The exact components and differences are proprietary.
- The RNAlater Stabilization Solution preserves RNA from degradation without lysing blood cells.
- Samples in RNAlater Stabilization Solution can be safely stored for up to 1 week at 25 degrees C, or up to 1 month at 4 degrees C.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.
Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.