I have some questions about Thermo's RNALAter Solution. Can it used for storage cells? Does anyone know whats the protocol? So, if I store my cells in this RNALater solution, can I still extract my RNA later and do a PCR reaction?
Someone used this RNALater solution ? Please give me some advice.
Actually, RNALater is just a near-saturated Ammonium Sulfate solution buffered with Sodium Citrate, and with some EDTA for metal chelation. Essentially, it causes protein aggregation (mostly reversible) when added to a tissue or cell suspension. This is why you need to incubate your sample for a while at 4 oC after mixing it with RNALater: it allows the reagent to permeate tissues/cells while their interior is still liquid. High ionic strength and the presence of EDTA add to ribonuclease inhibition. RNAs will aggregate too, this is why you have to remove as much of the reagent as possible before you start your RNA isolation procedure. When working with bacterial cells, I usually spin them down, discard the RNALater, then wash once with 0.3 M NaCl in TE buffer, finally resuspend in TE buffer and continue with Trizol extraction.
You can make RNALater-like reagent yourself, the composition is as follows:
70% (w/v) Ammonium Sulfate
25 mM Sodium Citrate
10 mM EDTA
Adjust pH to 5.2 with H2SO4
(see discussion and protocols here: https://www.researchgate.net/post/Anybody_with_homemade_RNAlater_experience)
Yes, you can store your cells in it. Actually, RNALater has been designed exactly for cells or tissues storage for some time until you have to extract RNA from them.
Yes, you can. Do read the following link regarding all your queries. The link is very useful.
https://www.thermofisher.com/in/en/home/references/ambion-tech-support/rna-isolation/tech-notes/rnalater-faqs.html
SD
Dear sherry
Yes you can perform both PCR and RNA extraction using this solution. The gradient I guest is trehalose a kind of carbohydrate however companies hide it. You should know that the samples are not siutable for Protein investigation and WB tests. It is very important the ratio of samples and solution regard as 1/5. It is also important to drop and left the tube in -4C for 24 h and then store in -70 for long time.
Ambion is a famous brand I suggest you.
Best regards
Actually, RNALater is just a near-saturated Ammonium Sulfate solution buffered with Sodium Citrate, and with some EDTA for metal chelation. Essentially, it causes protein aggregation (mostly reversible) when added to a tissue or cell suspension. This is why you need to incubate your sample for a while at 4 oC after mixing it with RNALater: it allows the reagent to permeate tissues/cells while their interior is still liquid. High ionic strength and the presence of EDTA add to ribonuclease inhibition. RNAs will aggregate too, this is why you have to remove as much of the reagent as possible before you start your RNA isolation procedure. When working with bacterial cells, I usually spin them down, discard the RNALater, then wash once with 0.3 M NaCl in TE buffer, finally resuspend in TE buffer and continue with Trizol extraction.
You can make RNALater-like reagent yourself, the composition is as follows:
70% (w/v) Ammonium Sulfate
25 mM Sodium Citrate
10 mM EDTA
Adjust pH to 5.2 with H2SO4
(see discussion and protocols here: https://www.researchgate.net/post/Anybody_with_homemade_RNAlater_experience)
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